@article{wulff_hickner_watson_denning_belikoff_scott_2024, title={Antennal transcriptome analysis reveals sensory receptors potentially associated with host detection in the livestock pest Lucilia cuprina}, volume={17}, ISSN={["1756-3305"]}, DOI={10.1186/s13071-024-06391-6}, abstractNote={Abstract Background Lucilia cuprina (Wiedemann, 1830) (Diptera: Calliphoridae) is the main causative agent of flystrike of sheep in Australia and New Zealand. Female flies lay eggs in an open wound or natural orifice, and the developing larvae eat the host’s tissues, a condition called myiasis. To improve our understanding of host-seeking behavior, we quantified gene expression in male and female antennae based on their behavior. Methods A spatial olfactometer was used to evaluate the olfactory response of L. cuprina mated males and gravid females to fresh or rotting beef. Antennal RNA-Seq analysis was used to identify sensory receptors differentially expressed between groups. Results Lucilia cuprina females were more attracted to rotten compared to fresh beef (> fivefold increase). However, males and some females did not respond to either type of beef. RNA-Seq analysis was performed on antennae dissected from attracted females, non-attracted females and males. Transcripts encoding sensory receptors from 11 gene families were identified above a threshold (≥ 5 transcript per million) including 49 ATP-binding cassette transporters (ABCs), two ammonium transporters (AMTs), 37 odorant receptors (ORs), 16 ionotropic receptors (IRs), 5 gustatory receptors (GRs), 22 odorant-binding proteins (OBPs), 9 CD36-sensory neuron membrane proteins (CD36/SNMPs), 4 chemosensory proteins (CSPs), 4 myeloid lipid-recognition (ML) and Niemann-Pick C2 disease proteins (ML/NPC2), 2 pickpocket receptors (PPKs) and 3 transient receptor potential channels (TRPs). Differential expression analyses identified sex-biased sensory receptors. Conclusions We identified sensory receptors that were differentially expressed between the antennae of both sexes and hence may be associated with host detection by female flies. The most promising for future investigations were as follows: an odorant receptor (LcupOR46) which is female-biased in L. cuprina and Cochliomyia hominivorax Coquerel, 1858; an ABC transporter (ABC G23.1) that was the sole sensory receptor upregulated in the antennae of females attracted to rotting beef compared to non-attracted females; a female-biased ammonia transporter (AMT_Rh50), which was previously associated with ammonium detection in Drosophila melanogaster Meigen, 1830. This is the first report suggesting a possible role for ABC transporters in L. cuprina olfaction and potentially in other insects. Graphical Abstract}, number={1}, journal={PARASITES & VECTORS}, author={Wulff, Juan P. and Hickner, Paul V. and Watson, David W. and Denning, Steven S. and Belikoff, Esther J. and Scott, Maxwell J.}, year={2024}, month={Jul} } @article{arp_williamson_vasquez_quintero_lowman_sagel_scott_2024, title={Doxycycline is a viable alternative to tetracycline for use in insect Tet-Off transgenic sexing systems, as assessed in the blowflies Cochliomyia hominivorax and Lucilia cuprina (Diptera: Calliphoridae)}, volume={2}, ISSN={["1938-291X"]}, url={https://doi.org/10.1093/jee/toae023}, DOI={10.1093/jee/toae023}, abstractNote={Abstract Transgenic insect strains with tetracycline repressible (Tet-Off) female-lethal genes provide significant advantages over traditional sterile insect techniques for insect population control, such as reduced diet and labor costs and more efficient population suppression. Tet-Off systems are suppressed by tetracycline-class antibiotics, most commonly tetracycline (Tc) or doxycycline (Dox), allowing for equal sex ratio colonies of transgenic insects when reared with Tc or Dox and male-only generations in their absence. Dox is a more stable molecule and has increased uptake than Tc, which could be advantageous in some insect mass-rearing systems. Here, we evaluated the suitability of Dox for rearing Tet-Off female-lethal strains of Australian sheep blowfly, Lucilia cuprina (Wiedemann, 1830) (Diptera: Calliphoridae), and New World screwworm, Cochliomyia hominivorax (Coquerel, 1858) (Diptera: Calliphoridae), and the effects of dosage on strain performance. For both species, colonies were able to be maintained with mixed-sex ratios at much lower dosages of Dox than Tc. Biological yields of C. hominivorax on either antibiotic were not significantly different. Reduction of Dox dosages in C. hominivorax diet did not affect biological performance, though rearing with 10 or 25 μg/mL was more productive than 50 μg/mL. Additionally, C. hominivorax mating performance and longevity were equal on all Dox dosages. Overall, Dox was a suitable antibiotic for mass-rearing Tet-Off female-lethal L. cuprina and C. hominivorax and was functional at much lower dosages than Tc.}, journal={JOURNAL OF ECONOMIC ENTOMOLOGY}, author={Arp, Alex P. and Williamson, Megan E. and Vasquez, Mario and Quintero, Gladys and Lowman, Aidamalia Vargas and Sagel, Agustin and Scott, Maxwell J.}, editor={Gerry, AlecEditor}, year={2024}, month={Feb} } @article{wulff_laminack_scott_2024, title={Genetic and behavioral analyses suggest that larval and adult stages ofLucilia cuprinaemploy different sensory systems to detect rotten beef}, url={https://doi.org/10.1101/2024.12.20.629795}, DOI={10.1101/2024.12.20.629795}, abstractNote={Abstract The blowfly Lucilia cuprina is a destructive parasite of sheep that causes flystrike or myiasis. Larvae consume the animal’s living flesh, producing large wounds that can lead to death. Growing resistance to conventional control methods has prompted the analysis of alternative strategies. RNA-Seq analysis of whole larvae at different stages and third instar head and gut tissues, suggested that odorant (OR), gustatory, ionotropic and pickpocke t receptors may not play a central role in the L. cuprina larval sensory signaling and digestive systems. Rather, ABC transporters were highly enriched in head and gut RNA, and OBPs only in the head. To confirm ORs are not essential for larval detection of rotten beef, diet-choice assays were performed including larvae and adults homozygous for a null mutation in the odorant coreceptor ( LcupOrco ) gene. While the attraction of adult females to rotten beef was fully disrupted, LcupOrco mutant larvae showed no change in diet preference.}, author={Wulff, Juan P. and Laminack, Rachel K. and Scott, Maxwell J.}, year={2024}, month={Dec} } @article{belikoff_davis_williamson_britt_scott_2024, title={Identification of a gene promoter active in Lucilia sericata larval salivary glands using a rapid transient expression assay}, volume={173}, ISSN={["1879-0240"]}, url={https://doi.org/10.1016/j.ibmb.2024.104163}, DOI={10.1016/j.ibmb.2024.104163}, journal={INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY}, author={Belikoff, Esther J. and Davis, Rebecca J. and Williamson, Megan E. and Britt, John W. and Scott, Maxwell J.}, year={2024}, month={Oct} } @article{yadav_butler_yamamoto_patil_lloyd_scott_2023, title={CRISPR/Cas9-based split homing gene drive targeting doublesex for population suppression of the global fruit pest Drosophila suzukii}, volume={120}, ISSN={0027-8424 1091-6490}, url={http://dx.doi.org/10.1073/pnas.2301525120}, DOI={10.1073/pnas.2301525120}, abstractNote={ Genetic-based methods offer environmentally friendly species-specific approaches for control of insect pests. One method, CRISPR homing gene drive that target genes essential for development, could provide very efficient and cost-effective control. While significant progress has been made in developing homing gene drives for mosquito disease vectors, little progress has been made with agricultural insect pests. Here, we report the development and evaluation of split homing drives that target the doublesex ( dsx ) gene in Drosophila suzukii , an invasive pest of soft-skinned fruits. The drive component, consisting of dsx single guide RNA and DsRed genes, was introduced into the female-specific exon of dsx , which is essential for function in females but not males. However, in most strains, hemizygous females were sterile and produced the male dsx transcript. With a modified homing drive that included an optimal splice acceptor site, hemizygous females from each of the four independent lines were fertile. High transmission rates of the DsRed gene (94 to 99%) were observed with a line that expressed Cas9 with two nuclear localization sequences from the D. suzukii nanos promoter. Mutant alleles of dsx with small in-frame deletions near the Cas9 cut site were not functional and thus would not provide resistance to drive. Finally, mathematical modeling showed that the strains could be used for suppression of lab cage populations of D. suzukii with repeated releases at relatively low release ratios (1:4). Our results indicate that the split CRISPR homing gene drive strains could potentially provide an effective means for control of D. suzukii populations. }, number={25}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Yadav, Amarish K. and Butler, Cole and Yamamoto, Akihiko and Patil, Anandrao A. and Lloyd, Alun L. and Scott, Maxwell J.}, year={2023}, month={Jun} } @article{patil_klobasa_espinoza‐rivera_baars_lorenzen_scott_2023, title={Development of transgenic corn planthopper Peregrinus maidis}, volume={32}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1111/imb.12836}, DOI={10.1111/imb.12836}, abstractNote={AbstractThe corn planthopper, Peregrinus maidis, is a vector of several maize viruses and is consequently a significant agricultural pest in many tropical and subtropical regions. As P. maidis has developed resistance to insecticides, the aim of this study was to develop transgenic P. maidis strains that could be used for future genetic biocontrol programs. To facilitate the identification of transgenic P. maidis, we isolated and characterized the promoters for the P. maidis ubiquitin‐like and profilin genes. Transient expression assays with P. maidis embryos showed that both promoters were active. Transgenic lines were established using piggyBac vectors and fluorescent protein marker genes. The lines carried an auto‐regulated tetracycline transactivator (tTA) gene, which has been widely used to establish conditional lethal strains in other insect species. The transgenic lines showed low levels of tTA expression but were viable on diet with or without doxycycline, which inhibits the binding of tTA to DNA. We discuss possible modifications to the tTA overexpression system that could lead to the successful development of conditional lethal strains. To our knowledge, this is the first report of a transgenic Hemiptera. The approach we have taken could potentially be applied to other Hemiptera and, for P. maidis, the technology will facilitate future functional genomics studies.}, number={4}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Patil, Anandrao A. and Klobasa, William and Espinoza‐Rivera, Dina and Baars, Oliver and Lorenzen, Marcé D. and Scott, Maxwell J.}, year={2023}, month={Mar}, pages={363–375} } @article{yadav_asokan_yamamoto_patil_scott_2023, title={Expansion of the genetic toolbox for manipulation of the global crop pest Drosophila suzukii: Isolation and assessment of eye colour mutant strains}, volume={10}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1111/imb.12879}, DOI={10.1111/imb.12879}, abstractNote={AbstractDrosophila suzukii (Matsumura) (Diptera: Drosophilidae), commonly called spotted wing Drosophila, is an important agricultural pest recognised worldwide. D. suzukii is a pest of soft‐skinned fruits as females can lay eggs in ripening fruit before harvest. While strains for genetic biocontrol of D. suzukii have been made, the development of transgenic D. suzukii strains and their further screening remain a challenge partly due to the lack of phenotypically trackable genetic‐markers, such as those widely used with the model genetic organism D. melanogaster. Here, we have used CRISPR/Cas9 to introduce heritable mutations in the eye colour genes white, cinnabar and sepia, which are located on the X, second and third chromosomes, respectively. Strains were obtained, which were homozygous for a single mutation. Genotyping of the established strains showed insertion and/or deletions (indels) at the targeted sites. A strain homozygous for mutations in cinnabar and sepia showed a pale‐yellow eye colour at eclosion but darkened to a sepia colour after a week. The fecundity and fertility of some of the cinnabar and sepia strains were comparable with the wild type. Although white mutant males were previously reported to be sterile, we found that sterility is not fully penetrant and we have been able to maintain white‐eyed strains for over a year. The cinnabar, sepia and white mutant strains developed in this study should facilitate future genetic studies in D. suzukii and the development of strains for genetic control of this pest.}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Yadav, Amarish K. and Asokan, Ramasamy and Yamamoto, Akihiko and Patil, Anandrao A. and Scott, Maxwell J.}, year={2023}, month={Oct} } @article{novas_basika_williamson_fresia_menchaca_scott_2023, title={Identification and functional analysis of Cochliomyia hominivoraxU6 gene promoters}, volume={32}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1111/imb.12875}, DOI={10.1111/imb.12875}, abstractNote={AbstractThe New World screwworm, Cochliomyia hominivorax, is an obligate parasite, which is a major pest of livestock. While the sterile insect technique was used very successfully to eradicate C. hominivorax from North and Central America, more cost‐effective genetic methods will likely be needed in South America. The recent development of CRISPR/Cas9‐based genetic approaches, such as homing gene drive, could provide a very efficient means for the suppression of C. hominivorax populations. One component of a drive system is the guide RNA(s) driven by a U6 gene promoter. Here, we have developed an in vivo assay to evaluate the activity of the promoters from seven C. hominivorax U6 genes. Embryos from the related blowfly Lucilia cuprina were injected with plasmid DNA containing a U6‐promoter‐guide RNA construct and a source of Cas9, either protein or plasmid DNA. Activity was assessed by the number of site‐specific mutations in the targeted gene in hatched larvae. One promoter, Chom U6_b, showed the highest activity. These U6 gene promoters could be used to build CRISPR/Cas9‐based genetic systems for the control of C. hominivorax.}, number={6}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Novas, Rossina and Basika, Tatiana and Williamson, Megan E. and Fresia, Pablo and Menchaca, Alejo and Scott, Maxwell J.}, year={2023}, month={Sep}, pages={716–724} } @article{tandonnet_krsticevic_basika_papathanos_torres_scott_2023, title={A chromosomal-scale reference genome of the New World Screwworm,Cochliomyia hominivorax}, volume={30}, ISSN={1340-2838 1756-1663}, url={http://dx.doi.org/10.1093/dnares/dsac042}, DOI={10.1093/dnares/dsac042}, abstractNote={AbstractThe New World Screwworm, Cochliomyia hominivorax (Calliphoridae), is the most important myiasis-causing species in America. Screwworm myiasis is a zoonosis that can cause severe lesions in livestock, domesticated and wild animals, and occasionally in people. Beyond the sanitary problems associated with this species, these infestations negatively impact economic sectors, such as the cattle industry. Here, we present a chromosome-scale assembly of C. hominivorax’s genome, organized in 6 chromosome-length and 515 unplaced scaffolds spanning 534 Mb. There was a clear correspondence between the D. melanogaster linkage groups A–E and the chromosomal-scale scaffolds. Chromosome quotient (CQ) analysis identified a single scaffold from the X chromosome that contains most of the orthologs of genes that are on the D. melanogaster fourth chromosome (linkage group F or dot chromosome). CQ analysis also identified potential X and Y unplaced scaffolds and genes. Y-linkage for selected regions was confirmed by PCR with male and female DNA. Some of the long chromosome-scale scaffolds include Y-linked sequences, suggesting misassembly of these regions. These resources will provide a basis for future studies aiming at understanding the biology and evolution of this devastating obligate parasite.}, number={1}, journal={DNA Research}, publisher={Oxford University Press (OUP)}, author={Tandonnet, Sophie and Krsticevic, Flavia and Basika, Tatiana and Papathanos, Philippos A and Torres, Tatiana T and Scott, Maxwell J}, year={2023}, month={Feb} } @misc{paulo_williamson_scott_2022, title={CRISPR/Cas9 Genome Editing in the New World Screwworm and Australian Sheep Blowfly}, ISBN={9781071623008 9781071623015}, ISSN={1064-3745 1940-6029}, url={http://dx.doi.org/10.1007/978-1-0716-2301-5_10}, DOI={10.1007/978-1-0716-2301-5_10}, abstractNote={Blowflies are of interest for medical applications (maggot therapy), forensic investigations, and for evolutionary developmental studies such as the evolution of parasitism. It is because of the latter that some blowflies such as the New World screwworm and the Australian sheep blowfly are considered major economic pests of livestock. Due to their importance, annotated assembled genomes for several species are now available. Here, we present a detailed guide for using the Streptococcus pyogenes Cas9 RNA-guided nuclease to efficiently generate both knockout and knock-in mutations in screwworm and sheep blowfly. These methods should accelerate genetic investigations in these and other closely related species and lead to a better understanding of the roles of selected genes in blowfly development and behavior.}, journal={Methods in Molecular Biology}, publisher={Springer US}, author={Paulo, Daniel F. and Williamson, Megan E. and Scott, Maxwell J.}, year={2022}, pages={173–201} } @article{yamamoto_yadav_scott_2022, title={Evaluation of Additional Drosophila suzukii Male-Only Strains Generated Through Remobilization of an FL19 Transgene}, volume={10}, ISSN={["2296-4185"]}, url={https://europepmc.org/articles/PMC8965018}, DOI={10.3389/fbioe.2022.829620}, abstractNote={Drosophila suzukii (D. suzukii) (Matsumura, 1931; Diptera: Drosophilidae), also known as spotted wing Drosophila, is a worldwide pest of fruits with soft skins such as blueberries and cherries. Originally from Asia, D. suzukii is now present in the Americas and Europe and has become a significant economic pest. Growers largely rely on insecticides for the control of D. suzukii. Genetic strategies offer a species-specific environmentally friendly way for suppression of D. suzukii populations. We previously developed a transgenic strain of D. suzukii that produced only males on a diet that did not contain tetracycline. The strain carried a single copy of the FL19 construct on chromosome 3. Repeated releases of an excess of FL19 males led to suppression of D. suzukii populations in laboratory cage trials. Females died as a consequence of overexpression of the tetracycline transactivator (tTA) and tTA-activated expression of the head involution defective proapoptotic gene. The aim of this study was to generate additional male-only strains that carried two copies of the FL19 transgene through crossing the original line with a piggyBac jumpstarter strain. Males that carried either two chromosome 3 or a singleX-linked transgene were identified through stronger expression of the red fluorescent protein marker gene. The brighter fluorescence of the X-linked lines was likely due to dosage compensation of the red fluorescent protein gene. In total, four X-linked lines and eleven lines with two copies on chromosome 3 were obtained, of which five were further examined. All but one of the strains produced only males on a diet without tetracycline. When crossed with wild type virgin females, all of the five two copy autosomal strains examined produced only males. However, the single copy X-linked lines did not show dominant female lethality. Five of the autosomal lines were further evaluated for productivity (egg to adult) and male competition. Based on these results, the most promising lines have been selected for future population suppression experiments with strains from different geographical locations.}, journal={FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY}, author={Yamamoto, Akihiko and Yadav, Amarish K. and Scott, Maxwell J.}, year={2022}, month={Mar} } @article{kandul_liu_buchman_shriner_corder_warsinger-pepe_yang_yadav_scott_marshall_et al._2022, title={Precision Guided Sterile Males Suppress Populations of an Invasive Crop Pest}, volume={1}, ISSN={2768-1572 2768-1556}, url={http://dx.doi.org/10.1089/genbio.2022.0019}, DOI={10.1089/genbio.2022.0019}, abstractNote={The Drosophila suzukii invasion of western countries has created an immense agricultural and economic threat to crop production. Despite many attempts to suppress its population, D. suzukii continues to destroy soft-flesh fruits. Precision guided sterile insect technique (pgSIT) utilizes the accuracy of programmable CRISPR gene targeting to generate sterilized males that can be deployed to suppress populations. Here, we generate pgSIT in D. suzukii and empirically and mathematically demonstrate that sterilized males are fit, competitive, and can eliminate populations of D. suzukii . Alto-gether, we describe an efficient way to generate sterile D. suzukii for release and safe effective population suppression.}, number={4}, journal={GEN Biotechnology}, publisher={Mary Ann Liebert Inc}, author={Kandul, Nikolay P. and Liu, Junru and Buchman, Anna and Shriner, Isaiah C. and Corder, Rodrigo M. and Warsinger-Pepe, Natalie and Yang, Ting and Yadav, Amarish K. and Scott, Maxwell J. and Marshall, John M. and et al.}, year={2022}, month={Aug}, pages={372–385} } @misc{scott_morrison_simmons_2022, title={Transgenic Approaches for Sterile Insect Control of Dipteran Livestock Pests and Lepidopteran Crop Pests}, ISBN={9781800621152 9781800621169}, url={http://dx.doi.org/10.1079/9781800621176.0017}, DOI={10.1079/9781800621176.0017}, journal={Transgenic Insects}, publisher={CABI}, author={Scott, Maxwell J. and Morrison, Neil I. and Simmons, Gregory S.}, year={2022}, month={Nov}, pages={340–358} } @book{benedict_scott_2022, place={Boston, MA}, title={Transgenic Insects}, ISBN={9781800621152 9781800621169}, url={http://dx.doi.org/10.1079/9781800621176.0000}, DOI={10.1079/9781800621176.0000}, journal={CABI}, publisher={CABI}, year={2022}, month={Nov} } @article{li_yamamoto_belikoff_berger_griffith_scott_2021, title={A conditional female lethal system for genetic suppression of the global fruit crop pest Drosophila suzukii}, volume={77}, ISSN={1526-498X 1526-4998}, url={http://dx.doi.org/10.1002/ps.6530}, DOI={10.1002/ps.6530}, abstractNote={AbstractBACKGROUNDDrosophila suzukii (Matsumura, 1931, Diptera: Drosophilidae) is a global pest of soft‐skinned fruits such as blueberries, cherries and raspberries. Also known as spotted‐wing drosophila, D. suzukii is native to Asia but is now widely distributed in the Americas and Europe, and presents a serious challenge for growers. Genetic control strategies offer an environmentally friendly approach for the control of D. suzukii.RESULTSIn this study, we developed transgenic strains of D. suzukii that carry dominant conditional female lethal transgenes. When raised in the absence of tetracycline, female D. suzukii die. We show that repeated releases of an excess of transgenic males can suppress D. suzukii populations in laboratory cage trials.CONCLUSIONOur data suggest that the transgenic strain could provide an effective approach for control of this invasive pest of soft‐skinned fruits.}, number={11}, journal={Pest Management Science}, publisher={Wiley}, author={Li, Fang and Yamamoto, Akihiko and Belikoff, Esther J and Berger, Amy and Griffith, Emily H and Scott, Maxwell J}, year={2021}, month={Jul}, pages={4915–4922} } @article{williamson_yan_scott_2021, title={Conditional knockdown of transformer in sheep blow fly suggests a role in repression of dosage compensation and potential for population suppression}, volume={17}, ISSN={1553-7404}, url={http://dx.doi.org/10.1371/journal.pgen.1009792}, DOI={10.1371/journal.pgen.1009792}, abstractNote={Thetransformer(tra) gene is essential for female development in many insect species, including the Australian sheep blow fly,Lucilia cuprina. Sex-specifictraRNA splicing is controlled bySex lethal(Sxl) inDrosophila melanogasterbut is auto-regulated inL.cuprina.Sxlalso represses X chromosome dosage compensation in femaleD.melanogaster. We have developed conditionalLctraRNAi knockdown strains using the tet-off system. Four strains did not produce females on diet without tetracycline and could potentially be used for genetic control ofL.cuprina. In one strain, which showed both maternal and zygotic tTA expression, most XX transformed males died at the pupal stage. RNAseq and qRT-PCR analyses of mid-stage pupae showed increased expression of X-linked genes in XX individuals. These results suggest thatLctrapromotes somatic sexual differentiation and inhibits X chromosome dosage compensation in femaleL.cuprina. However, XX flies homozygous for a loss-of-functionLctraknockin mutation were fully transformed and showed high pupal eclosion. Two of five X-linked genes examined showed a significant increase in mRNA levels in XX males. The stronger phenotype in the RNAi knockdown strain could indicate that maternalLctraexpression may be essential for initiation of dosage compensation suppression in female embryos.}, number={10}, journal={PLOS Genetics}, publisher={Public Library of Science (PLoS)}, author={Williamson, Megan E. and Yan, Ying and Scott, Maxwell J.}, editor={Palli, Subba ReddyEditor}, year={2021}, month={Oct}, pages={e1009792} } @article{li_yamamoto_belikoff_berger_griffith_scott_2021, title={Cover Image, Volume 77, Issue 11}, volume={77}, url={https://doi.org/10.1002/ps.6661}, DOI={10.1002/ps.6661}, abstractNote={The cover image is based on the Research Article A conditional female lethal system for genetic suppression of the global fruit crop pest Drosophila suzukii by Fang Li et al., https://doi.org/10.1002/ps.6530. Design Credit: Dr. Amarish Yadav and Dr. Akihiko Yamamoto.image}, number={11}, journal={Pest Management Science}, publisher={Wiley}, author={Li, Fang and Yamamoto, Akihiko and Belikoff, Esther J and Berger, Amy and Griffith, Emily H and Scott, Maxwell J}, year={2021}, month={Nov} } @article{paulo_junqueira_arp_vieira_ceballos_skoda_pérez-de-león_sagel_mcmillan_scott_et al._2021, title={Disruption of the odorant coreceptor Orco impairs foraging and host finding behaviors in the New World screwworm fly}, volume={11}, ISSN={2045-2322}, url={http://dx.doi.org/10.1038/s41598-021-90649-x}, DOI={10.1038/s41598-021-90649-x}, abstractNote={AbstractThe evolution of obligate ectoparasitism in blowflies (Diptera: Calliphoridae) has intrigued scientists for over a century, and surprisingly, the genetics underlying this lifestyle remain largely unknown. Blowflies use odors to locate food and oviposition sites; therefore, olfaction might have played a central role in niche specialization within the group. In insects, the coreceptor Orco is a required partner for all odorant receptors (ORs), a major gene family involved in olfactory-evoked behaviors. Hence, we characterized the Orco gene in the New World screwworm, Cochliomyia hominivorax, a blowfly that is an obligate ectoparasite of warm-blooded animals. In contrast, most of the closely related blowflies are scavengers that lay their eggs on dead animals. We show that the screwworm Orco orthologue (ChomOrco) is highly conserved within Diptera, showing signals of strong purifying selection. Expression of ChomOrco is broadly detectable in chemosensory appendages, and is related to morphological, developmental, and behavioral aspects of the screwworm biology. We used CRISPR/Cas9 to disrupt ChomOrco and evaluate the consequences of losing the OR function on screwworm behavior. In two-choice assays, Orco mutants displayed an impaired response to floral-like and animal host-associated odors, suggesting that OR-mediated olfaction is involved in foraging and host-seeking behaviors in C. hominivorax. These results broaden our understanding of the chemoreception basis of niche occupancy by blowflies.}, number={1}, journal={Scientific Reports}, publisher={Springer Science and Business Media LLC}, author={Paulo, Daniel F. and Junqueira, Ana C. M. and Arp, Alex P. and Vieira, André S. and Ceballos, Jorge and Skoda, Steven R. and Pérez-de-León, Adalberto A. and Sagel, Agustin and McMillan, William O. and Scott, Maxwell J. and et al.}, year={2021}, month={May} } @article{tait_mermer_stockton_lee_avosani_abrieux_anfora_beers_biondi_burrack_et al._2021, title={Drosophila suzukii (Diptera: Drosophilidae): A Decade of Research Towards a Sustainable Integrated Pest Management Program}, volume={114}, ISSN={0022-0493 1938-291X}, url={http://dx.doi.org/10.1093/jee/toab158}, DOI={10.1093/jee/toab158}, abstractNote={Abstract Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) also known as spotted-wing drosophila (SWD), is a pest native to Southeast Asia. In the last few decades, the pest has expanded its range to affect all major European and American fruit production regions. SWD is a highly adaptive insect that is able to disperse, survive, and flourish under a range of environmental conditions. Infestation by SWD generates both direct and indirect economic impacts through yield losses, shorter shelf life of infested fruit, and increased production costs. Fresh markets, frozen berries, and fruit export programs have been impacted by the pest due to zero tolerance for fruit infestation. As SWD control programs rely heavily on insecticides, exceedance of maximum residue levels (MRLs) has also resulted in crop rejections. The economic impact of SWD has been particularly severe for organic operations, mainly due to the limited availability of effective insecticides. Integrated pest management (IPM) of SWD could significantly reduce chemical inputs but would require substantial changes to horticultural management practices. This review evaluates the most promising methods studied as part of an IPM strategy against SWD across the world. For each of the considered techniques, the effectiveness, impact, sustainability, and stage of development are discussed.}, number={5}, journal={Journal of Economic Entomology}, publisher={Oxford University Press (OUP)}, author={Tait, Gabriella and Mermer, Serhan and Stockton, Dara and Lee, Jana and Avosani, Sabina and Abrieux, Antoine and Anfora, Gianfranco and Beers, Elizabeth and Biondi, Antonio and Burrack, Hannah and et al.}, editor={Brewer, MichaelEditor}, year={2021}, month={Sep}, pages={1950–1974} } @article{kandul_belikoff_liu_buchman_li_yamamoto_yang_shriner_scott_akbari_2021, title={Genetically Encoded CRISPR Components Yield Efficient Gene Editing in the Invasive Pest Drosophila suzukii}, volume={4}, ISSN={2573-1599 2573-1602}, url={http://dx.doi.org/10.1089/crispr.2021.0032}, DOI={10.1089/crispr.2021.0032}, abstractNote={Originally from Asia, Drosophila suzukii Matsumura is a global pest of economically important soft-skinned fruits. Also commonly known as spotted wing drosophila, it is largely controlled through repeated applications of broad-spectrum insecticides by which resistance has been observed in the field. There is a pressing need for a better understanding of D. suzukii biology and for developing alternative environmentally friendly methods of control. The RNA-guided Cas9 nuclease has revolutionized functional genomics and is an integral component of several recently developed genetic strategies for population control of insects. Here, we describe genetically modified strains that encode three different terminators and four different promoters to express Cas9 robustly in both the soma and/or germline of D. suzukii. The Cas9 strains were rigorously evaluated through genetic crossing to transgenic strains that encode single-guide RNAs targeting the conserved X-linked yellow body and white eye genes. We find that several Cas9/gRNA strains display remarkably high editing capacity. Going forward, these tools will be instrumental for evaluating gene function in D. suzukii and may even provide tools useful for the development of new genetic strategies for control of this invasive species.}, number={5}, journal={The CRISPR Journal}, publisher={Mary Ann Liebert Inc}, author={Kandul, Nikolay P. and Belikoff, Esther J. and Liu, Junru and Buchman, Anna and Li, Fang and Yamamoto, Akihiko and Yang, Ting and Shriner, Isaiah and Scott, Maxwell J. and Akbari, Omar S.}, year={2021}, month={Oct}, pages={739–751} } @article{davis_belikoff_dickey_scholl_benoit_scott_2021, title={Genome and transcriptome sequencing of the green bottle fly, Lucilia sericata, reveals underlying factors of sheep flystrike and maggot debridement therapy}, volume={113}, ISSN={0888-7543}, url={http://dx.doi.org/10.1016/j.ygeno.2021.10.003}, DOI={10.1016/j.ygeno.2021.10.003}, abstractNote={The common green bottle blow fly Lucilia sericata (family, Calliphoridae) is widely used for maggot debridement therapy, which involves the application of sterile maggots to wounds. The larval excretions and secretions are important for consuming necrotic tissue and inhibiting bacterial growth in wounds of patients. Lucilia sericata is also of importance as a pest of sheep and in forensic studies to estimate a postmortem interval. Here we report the assembly of a 565.3 Mb genome from long read PacBio DNA sequencing of genomic DNA. The genome contains 14,704 predicted protein coding genes and 1709 non-coding genes. Targeted annotation and transcriptional analyses identified genes that are highly expressed in the larval salivary glands (secretions) and Malpighian tubules (excretions) under normal growth conditions and following heat stress. The genomic resources will underpin future genetic studies and in development of engineered strains for genetic control of L. sericata and for biotechnology-enhanced maggot therapy.}, number={6}, journal={Genomics}, publisher={Elsevier BV}, author={Davis, Rebecca J. and Belikoff, Esther J. and Dickey, Allison N. and Scholl, Elizabeth H. and Benoit, Joshua B. and Scott, Maxwell J.}, year={2021}, month={Nov}, pages={3978–3988} } @article{scott_2021, title={Sex Determination and Dosage Compensation: femaleless Is the Link in Anopheles Mosquitoes}, volume={31}, ISSN={0960-9822}, url={http://dx.doi.org/10.1016/j.cub.2021.01.078}, DOI={10.1016/j.cub.2021.01.078}, abstractNote={A new study finds that the femaleless gene is essential for sexual development and repression of X-chromosome dosage compensation in the malaria vector Anopheles gambiae. This could provide the basis for a new genetic approach to control this pest.}, number={5}, journal={Current Biology}, publisher={Elsevier BV}, author={Scott, Max}, year={2021}, month={Mar}, pages={R260–R263} } @article{webster_scott_2021, title={TheAedes aegypti(Diptera: Culicidae)hsp83Gene Promoter Drives Strong Ubiquitous DsRed and ZsGreen Marker Expression in Transgenic Mosquitoes}, volume={58}, ISSN={0022-2585 1938-2928}, url={http://dx.doi.org/10.1093/jme/tjab128}, DOI={10.1093/jme/tjab128}, abstractNote={AbstractTransgenic strains of the mosquito disease vector Aedes aegypti (L.) are being developed for population suppression or modification. Transgenic mosquitoes are identified using fluorescent protein genes. Here we describe DsRed and ZsGreen marker genes driven by the constitutive Ae. aegypti heat shock protein 83 (hsp83) promoter in transgenic mosquitoes. Transgenic larvae and pupae show strong full body expression of the red and green fluorescent proteins. This greatly assists in screening for transgenic individuals while making new or maintaining already established lines. Transient marker gene expression after embryo microinjection was readily visible in developing larvae allowing the separation of individuals that are more likely to produce transgenic offspring. The strongly expressed marker genes developed in this study should facilitate the detection of transgenic Ae. aegypti larvae or pupae in the field.}, number={6}, journal={Journal of Medical Entomology}, publisher={Oxford University Press (OUP)}, author={Webster, Sophia H and Scott, Maxwell J}, editor={Slotman, Michel AEditor}, year={2021}, month={Jul}, pages={2533–2537} } @article{concha_yan_arp_quilarque_sagel_de león_mcmillan_skoda_scott_2020, title={An early female lethal system of the New World screwworm, Cochliomyia hominivorax, for biotechnology-enhanced SIT}, volume={21}, ISSN={1471-2156}, url={http://dx.doi.org/10.1186/s12863-020-00948-x}, DOI={10.1186/s12863-020-00948-x}, abstractNote={AbstractBackgroundThe New World Screwworm fly (NWS),Cochliomyia hominivorax, is an ectoparasite of warm-blooded animals and a major pest of livestock in parts of South America and the Caribbean where it remains endemic. In North and Central America it was eradicated using the Sterile Insect Technique (SIT). A control program is managed cooperatively between the governments of the United States and Panama to prevent the northward spread of NWS from infested countries in South America. This is accomplished by maintaining a permanent barrier through the release of millions of sterile male and female flies in the border between Panama and Colombia. Our research team demonstrated the utility of biotechnology-enhanced approaches for SIT by developing a male-only strain of the NWS. The strain carried a single component tetracycline repressible female lethal system where females died at late larval/pupal stages. The control program can be further improved by removing females during embryonic development as larval diet costs are significant.ResultsThe strains developed carry a two-component system consisting of theLucilia sericata bottleneckgene promoter driving expression of the tTA gene and a tTA-regulatedLshidproapoptotic effector gene. Insertion of the sex-specifically spliced intron from theC. hominivorax transformergene within theLshidgene ensures that only females die when insects are reared in the absence of tetracycline. In several double homozygous two-component strains and in one “All-in-one” strain that had both components in a single construct, female lethality occurred at the embryonic and/or first instar larval stages when raised on diet without tetracycline. Laboratory evaluation for phenotypes that are relevant for mass rearing in a production facility revealed that most strains had fitness characteristics similar to the wild type J06 strain that is currently reared for release in the permanent barrier. Testing of an “All in one” strain under mass rearing conditions showed that the strain maintained the fitness characteristics observed in small-scale rearing.ConclusionsThe early female lethal strains described here could be selected by the NWS Control Program for testing at large scale in the production facility to enhance the efficiency of the NWS eradication program.}, number={S2}, journal={BMC Genetics}, publisher={Springer Science and Business Media LLC}, author={Concha, Carolina and Yan, Ying and Arp, Alex and Quilarque, Evelin and Sagel, Agustin and de León, Adalberto Pérez and McMillan, W. Owen and Skoda, Steven and Scott, Maxwell J.}, year={2020}, month={Dec} } @article{yan_scott_2020, title={Building a transgenic sexing strain for genetic control of the Australian sheep blow fly Lucilia cuprina using two lethal effectors}, volume={21}, ISSN={1471-2156}, url={http://dx.doi.org/10.1186/s12863-020-00947-y}, DOI={10.1186/s12863-020-00947-y}, abstractNote={Abstract Background The sterile insect technique (SIT) has been successfully used in many pest management programs worldwide. Some SIT programs release both sexes due to the lack of genetic sexing strains or efficient sex separation methods but sterile females are ineffective control agents. Transgenic sexing strains (TSS) using the tetracycline-off control system have been developed in a variety of insect pests, from which females die by either of two commonly used lethal effectors: overexpression of the transcription factor tetracycline transactivator (tTA) or ectopic expression of a proapoptotic gene, such as head involution defective (hid). The lethality from tTA overexpression is thought to be due to “transcriptional squelching”, while hid causes lethality by induction of apoptosis. This study aims to create and characterize a TSS of Lucilia cuprina, which is a major pest of sheep, by combining both lethal effectors in a single transgenic strain. Results Here a stable TSS of L. cuprina (DH6) that carries two lethal effectors was successfully generated, by crossing FL3#2 which carries a female-specific tTA overexpression cassette, with EF1#12 which carries a tTA-regulated LshidAla2 cassette. Females with one copy of the FL3#2 transgene are viable but up to 99.8% of homozygous females die at the pupal stage when raised on diet that lacks tetracycline. Additionally, the female lethality of FL3#2 was partially repressed by supplying tetracycline to the parental generation. With an additional LshidAla2 effector, the female lethality of DH6 is 100% dominant and cannot be repressed by maternal tetracycline. DH6 females die at the late-larval stage. Several fitness parameters important for mass rearing such as hatching rate, adult emergence and sex ratio were comparable to those of the wild type strain. Conclusions Compared to the parental FL3#2 strain, the DH6 strain shows stronger female lethality and lethality occurs at an earlier stage of development. The combination of two tTA-dependent lethal effectors could improve strain stability under mass rearing and could reduce the risk of resistance in the field if fertile males are released. Our approach could be easily adapted for other pest species for an efficient, safe and sustainable genetic control program. }, number={S2}, journal={BMC Genetics}, publisher={Springer Science and Business Media LLC}, author={Yan, Ying and Scott, Maxwell J.}, year={2020}, month={Dec} } @misc{jaffri_yan_scott_schetelig_2020, title={Conditional Expression Systems for Drosophila suzukii Pest Control}, ISBN={9783030626914 9783030626921}, url={http://dx.doi.org/10.1007/978-3-030-62692-1_10}, DOI={10.1007/978-3-030-62692-1_10}, journal={Drosophila suzukii Management}, publisher={Springer International Publishing}, author={Jaffri, Syeda A. and Yan, Ying and Scott, Maxwell J. and Schetelig, Marc F.}, year={2020}, pages={195–215} } @article{long_alphey_annas_bloss_campbell_champer_chen_choudhary_church_collins_et al._2020, title={Core commitments for field trials of gene drive organisms}, volume={370}, ISSN={0036-8075 1095-9203}, url={http://dx.doi.org/10.1126/science.abd1908}, DOI={10.1126/science.abd1908}, abstractNote={We must ensure that trials are scientifically, politically, and socially robust, publicly accountable, and widely transparent}, number={6523}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Long, Kanya C. and Alphey, Luke and Annas, George J. and Bloss, Cinnamon S. and Campbell, Karl J. and Champer, Jackson and Chen, Chun-Hong and Choudhary, Amit and Church, George M. and Collins, James P. and et al.}, year={2020}, month={Dec}, pages={1417–1419} } @article{webster_vella_scott_2020, title={Development and testing of a novel killer–rescue self-limiting gene drive system in Drosophila melanogaster}, volume={287}, ISSN={0962-8452 1471-2954}, url={http://dx.doi.org/10.1098/rspb.2019.2994}, DOI={10.1098/rspb.2019.2994}, abstractNote={ Here we report the development and testing of a novel self-limiting gene drive system, Killer–Rescue (K–R), in Drosophila melanogaster . This system is composed of an autoregulated Gal4 Killer (K) and a Gal4-activated Gal80 Rescue (R). Overexpression of Gal4 is lethal, but in the presence of R activation of Gal80 leads to much lower levels of Gal4 and rescue of lethality. We demonstrate that with a single 2 : 1 engineered to wild-type release, K drives R through the population and after nine generations, more than 98% of the population carry R and less than 2% of the population are wild-type flies. We discuss how this simple K–R gene drive system may be readily adapted for population replacement in a human health pest, Aedes aegypti , or for population suppression in an agricultural pest, Drosophila suzukii . }, number={1925}, journal={Proceedings of the Royal Society B: Biological Sciences}, publisher={The Royal Society}, author={Webster, Sophia H. and Vella, Michael R. and Scott, Maxwell J.}, year={2020}, month={Apr}, pages={20192994} } @article{knudsen_reid_barbour_bowes_duncan_philpott_potter_scott_2020, title={Genetic Variation and Potential for Resistance Development to the tTA Overexpression Lethal System in Insects}, volume={10}, ISSN={2160-1836}, url={http://dx.doi.org/10.1534/g3.120.400990}, DOI={10.1534/g3.120.400990}, abstractNote={AbstractRelease of insect pests carrying the dominant lethal tetracycline transactivator (tTA) overexpression system has been proposed as a means for population suppression. High levels of the tTA transcription factor are thought to be toxic due to either transcriptional squelching or interference with protein ubiquitination. Here we utilized the Drosophila melanogaster Genetic Reference Panel (DGRP) to examine the influence of genetic variation on the efficacy of a female-specific tTA overexpression system. The level of female lethality between DGRP lines varied from 11 to 97% with a broad sense heritability of 0.89. A genome-wide association analysis identified 192 allelic variants associated with high or low lethality (P < 10−5), although none were significant when corrected for multiple testing. 151 of the variants fell within 108 genes that were associated with several biological processes including transcription and protein ubiquitination. In four lines with high female lethality, tTA RNA levels were similar or higher than in the parental tTA overexpression strain. In two lines with low lethality, tTA levels were about two fold lower than in the parental strain. However, in two other lines with low lethality, tTA levels were similar or approximately 30% lower. RNAseq analysis identified genes that were up or downregulated in the four low female lethal lines compared to the four high lethal lines. For example, genes associated with RNA processing and rRNA maturation were significantly upregulated in low lethal lines. Our data suggest that standing genetic variation in an insect population could provide multiple mechanisms for resistance to the tTA overexpression system.}, number={4}, journal={G3 Genes|Genomes|Genetics}, publisher={Oxford University Press (OUP)}, author={Knudsen, Katherine E and Reid, William R and Barbour, Traci M and Bowes, Laci M and Duncan, Juliana and Philpott, Elaina and Potter, Samantha and Scott, Maxwell J}, year={2020}, month={Apr}, pages={1271–1281} } @article{scott_benoit_davis_bailey_varga_martinson_hickner_syed_cardoso_torres_et al._2020, title={Genomic analyses of a livestock pest, the New World screwworm, find potential targets for genetic control programs}, volume={3}, ISSN={2399-3642}, url={http://dx.doi.org/10.1038/s42003-020-01152-4}, DOI={10.1038/s42003-020-01152-4}, abstractNote={AbstractThe New World Screwworm fly, Cochliomyia hominivorax, is a major pest of livestock in South America and Caribbean. However, few genomic resources have been available for this species. A genome of 534 Mb was assembled from long read PacBio DNA sequencing of DNA from a highly inbred strain. Analysis of molecular evolution identified 40 genes that are likely under positive selection. Developmental RNA-seq analysis identified specific genes associated with each stage. We identify and analyze the expression of genes that are likely important for host-seeking behavior (chemosensory), development of larvae in open wounds in warm-blooded animals (heat shock protein, immune response) and for building transgenic strains for genetic control programs including gene drive (sex determination, germline). This study will underpin future experiments aimed at understanding the parasitic lifestyle of the screwworm fly and greatly facilitate future development of strains for efficient systems for genetic control of screwworm.}, number={1}, journal={Communications Biology}, publisher={Springer Science and Business Media LLC}, author={Scott, Maxwell J. and Benoit, Joshua B. and Davis, Rebecca J. and Bailey, Samuel T. and Varga, Virag and Martinson, Ellen O. and Hickner, Paul V. and Syed, Zainulabeuddin and Cardoso, Gisele A. and Torres, Tatiana T. and et al.}, year={2020}, month={Aug} } @article{hickner_mittapalli_subramoniam_sagel_watson_scott_arp_de león_syed_2020, title={Physiological and molecular correlates of the screwworm fly attraction to wound and animal odors}, volume={10}, ISBN={2045-2322}, ISSN={2045-2322}, url={http://dx.doi.org/10.1038/s41598-020-77541-w}, DOI={10.1038/s41598-020-77541-w}, abstractNote={AbstractThe screwworm fly,Cochliomyia hominivorax(Coquerel), was successfully eradicated from the United States by the sterile insect technique (SIT). However, recent detection of these flies in the Florida Keys, and increased risk of introductions to the other areas warrant novel tools for management of the flies. Surveillance, a key component of screwworm control programs, utilizes traps baited with rotting liver or a blend of synthetic chemicals such asswormlure-4. In this work, we evaluated the olfactory physiology of the screwworm fly and compared it with the non-obligate ectoparasitic secondary screwworm flies,C. macellaria,that invade necrotic wound and feed on dead tissue. These two species occur in geographically overlapping regions.C. macellaria, along with other blowflies such as the exoticC. megacephala, greatly outnumberC. hominivoraxin the existing monitoring traps. Olfactory responses toswormlure-4constituents between sex and mating status (mated vs unmated) in both species were recorded and compared. Overall, responses measured by the antennograms offered insights into the comparative olfactory physiology of the two fly species. We also present detailed analyses of the antennal transcriptome by RNA-Sequencing that reveal significant differences between male and female screwworm flies. The differential expression patterns were confirmed by quantitative PCR. Taken together, this integrated study provides insights into the physiological and molecular correlates of the screwworm’s attraction to wounds, and identifies molecular targets that will aid in the development of odorant-based fly management strategies.}, number={1}, journal={Scientific Reports}, publisher={Springer Science and Business Media LLC}, author={Hickner, Paul V. and Mittapalli, Omprakash and Subramoniam, Anjana and Sagel, Agustin and Watson, Wes and Scott, Maxwell J. and Arp, Alex P. and de León, Adalberto A. Pérez and Syed, Zainulabeuddin}, year={2020}, month={Nov} } @article{yan_williamson_scott_2020, title={Using Moderate Transgene Expression to Improve the Genetic Sexing System of the Australian Sheep Blow Fly Lucilia cuprina}, volume={11}, ISSN={2075-4450}, url={http://dx.doi.org/10.3390/insects11110797}, DOI={10.3390/insects11110797}, abstractNote={The sterile insect technique (SIT) is a promising strategy to control the Australian sheep blow fly Lucilia cuprina, a major pest of sheep. We have previously developed a transgenic embryonic sexing system (TESS) for this pest to facilitate the potential SIT application. TESS carry two transgenes, a tetracycline transactivator (tTA) driver and a tTA-activated pro-apoptotic effector. TESS females die at the embryonic stage unless tetracycline is supplied in the diet. However, undesired female sterility was observed in some TESS strains without tetracycline due to expression of tTA in ovaries. Here we investigate if TESS that combine transgenes with relatively low/moderate expression/activity improves the fertility of TESS females. tTA driver lines were evaluated for tTA expression by quantitative real time PCR and/or by crossing with a tTA-activated RFPex effector line. Fertility and lethality tests showed that a TESS strain containing a driver line with moderate tTA expression and an effector line showing moderate pro-apoptotic activity could recover the fertility of parental females and eliminated all female offspring at the embryonic stage. Consequently, such a strain could be further evaluated for an SIT program for L. cuprina, and such a “moderate strategy” could be considered for the TESS development in other pest species.}, number={11}, journal={Insects}, publisher={MDPI AG}, author={Yan, Ying and Williamson, Megan E. and Scott, Maxwell J.}, year={2020}, month={Nov}, pages={797} } @article{yan_williamson_davis_andere_picard_scott_2019, title={Improved transgenic sexing strains for genetic control of the Australian sheep blow fly Lucilia cuprina using embryo-specific gene promoters}, volume={295}, ISSN={1617-4615 1617-4623}, url={http://dx.doi.org/10.1007/s00438-019-01622-3}, DOI={10.1007/s00438-019-01622-3}, abstractNote={For genetic approaches for controlling insect pests such as the sterile insect technique (SIT), it is advantageous to release only males as females are ineffective as control agents and they consume about 50% of the diet. Here we developed tetracycline-repressible Lucilia cuprina transgenic strains in which adult females were fully fertile and viable on a diet that lacked tetracycline and all of their female offspring died at the embryo stage. The transgenic strains are an improvement over the strains we developed previously, which had the disadvantage that adult females on diet without tetracycline were sterile and died prematurely. This was possibly due to the low level expression of the effector gene in ovaries. In the strains developed in this study, the early promoters from L. cuprina nullo or Cochliomyia macellaria CG14427 genes were used to drive the tetracycline transactivator (tTA) expression in the early embryo. In the absence of tetracycline, tTA activates expression of the proapoptotic gene Lshid which contains a female-specific intron. Consequently, only females produce active HID protein and die at the embryo stage. Crossing the tTA-expressing driver lines with an RFPex reporter line confirmed that there was no expression of the effector gene in the ovary. These new embryonic L. cuprina transgenic sexing strains hold great promise for genetic control programs and the system reported here might also be transferable to other major calliphorid livestock pests such as the New World screwworm, Cochliomyia hominivorax.}, number={2}, journal={Molecular Genetics and Genomics}, publisher={Springer Science and Business Media LLC}, author={Yan, Ying and Williamson, Megan E. and Davis, Rebecca J. and Andere, Anne A. and Picard, Christine J. and Scott, Maxwell J.}, year={2019}, month={Nov}, pages={287–298} } @article{concha_wallbank_hanly_fenner_livraghi_rivera_paulo_arias_vargas_sanjeev_et al._2019, title={Interplay between Developmental Flexibility and Determinism in the Evolution of Mimetic Heliconius Wing Patterns}, volume={29}, ISSN={0960-9822}, url={http://dx.doi.org/10.1016/j.cub.2019.10.010}, DOI={10.1016/j.cub.2019.10.010}, abstractNote={To what extent can we predict how evolution occurs? Do genetic architectures and developmental processes canalize the evolution of similar outcomes in a predictable manner? Or do historical contingencies impose alternative pathways to answer the same challenge? Examples of Müllerian mimicry between distantly related butterfly species provide natural replicates of evolution, allowing us to test whether identical wing patterns followed parallel or novel trajectories. Here, we explore the role that the signaling ligand WntA plays in generating mimetic wing patterns in Heliconius butterflies, a group with extraordinary mimicry-related wing pattern diversity. The radiation is relatively young, and numerous cases of wing pattern mimicry have evolved within the last 2.5-4.5 Ma. WntA is an important target of natural selection and is one of four major effect loci that underlie much of the pattern variation in the group. We used CRISPR/Cas9 targeted mutagenesis to generate WntA-deficient wings in 12 species and a further 10 intraspecific variants, including three co-mimetic pairs. In all tested butterflies, WntA knockouts affect pattern broadly and cause a shift among every possible scale cell type. Interestingly, the co-mimics lacking WntA were very different, suggesting that the gene networks that pattern a wing have diverged considerably among different lineages. Thus, although natural selection channeled phenotypic convergence, divergent developmental contexts between the two major Heliconius lineages opened different developmental routes to evolve resemblance. Consequently, even under very deterministic evolutionary scenarios, our results underscore a surprising unpredictability in the developmental paths underlying convergence in a recent radiation.}, number={23}, journal={Current Biology}, publisher={Elsevier BV}, author={Concha, Carolina and Wallbank, Richard W.R. and Hanly, Joseph J. and Fenner, Jennifer and Livraghi, Luca and Rivera, Edgardo Santiago and Paulo, Daniel F. and Arias, Carlos and Vargas, Marta and Sanjeev, Manu and et al.}, year={2019}, month={Dec}, pages={3996–4009.e4} } @article{paulo_williamson_arp_li_sagel_skoda_sanchez-gallego_vasquez_quintero_pérez de león_et al._2019, title={Specific Gene Disruption in the Major Livestock PestsCochliomyia hominivoraxandLucilia cuprinaUsing CRISPR/Cas9}, volume={9}, ISSN={2160-1836}, url={http://dx.doi.org/10.1534/g3.119.400544}, DOI={10.1534/g3.119.400544}, abstractNote={AbstractCochliomyia hominivorax and Lucilia cuprina are major pests of livestock. Their larvae infest warm-blooded vertebrates and feed on host’s tissues, resulting in severe industry losses. As they are serious pests, considerable effort has been made to develop genomic resources and functional tools aiming to improve their management and control. Here, we report a significant addition to the pool of genome manipulation tools through the establishment of efficient CRISPR/Cas9 protocols for the generation of directed and inheritable modifications in the genome of these flies. Site-directed mutations were introduced in the C. hominivorax and L. cuprina yellow genes (ChY and LcY) producing lightly pigmented adults. High rates of somatic mosaicism were induced when embryos were injected with Cas9 ribonucleoprotein complexes (RNPs) pre-assembled with guide RNAs (sgRNAs) at high concentrations. Adult flies carrying disrupted yellow alleles lacked normal pigmentation (brown body phenotype) and efficiently transmitted the mutated alleles to the subsequent generation, allowing the rapid creation of homozygous strains for reverse genetics of candidate loci. We next used our established CRISPR protocol to disrupt the C. hominivorax transformer gene (Chtra). Surviving females carrying mutations in the Chtra locus developed mosaic phenotypes of transformed ovipositors with characteristics of male genitalia while exhibiting abnormal reproductive tissues. The CRISPR protocol described here is a significant improvement on the existing toolkit of molecular methods in calliphorids. Our results also suggest that Cas9-based systems targeting Chtra and Lctra could be an effective means for controlling natural populations of these important pests.}, number={9}, journal={G3 Genes|Genomes|Genetics}, publisher={Oxford University Press (OUP)}, author={Paulo, Daniel F and Williamson, Megan E and Arp, Alex P and Li, Fang and Sagel, Agustin and Skoda, Steven R and Sanchez-Gallego, Joel and Vasquez, Mario and Quintero, Gladys and Pérez de León, Adalberto A and et al.}, year={2019}, month={Sep}, pages={3045–3055} } @article{davis_belikoff_scott_2018, title={Towards next generation maggot debridement therapy: Transgenic Lucilia sericata larvae that produce and secrete a human growth factor}, volume={26}, number={1}, journal={Wound Repair and Regeneration}, author={Davis, R. J. and Belikoff, E. J. and Scott, M. J.}, year={2018}, pages={A27–27} } @article{davis_belikoff_scholl_li_scott_2018, title={no blokes Is Essential for Male Viability and X Chromosome Gene Expression in the Australian Sheep Blowfly}, volume={28}, ISSN={0960-9822}, url={http://dx.doi.org/10.1016/j.cub.2018.05.005}, DOI={10.1016/j.cub.2018.05.005}, abstractNote={It has been hypothesized that the Drosophila 4 th  chromosome is derived from an ancient X chromosome [1]. In the Australian sheep blowfly, Lucilia cuprina, the heterochromatic X chromosome contains few active genes and orthologs of Drosophila X-linked genes are autosomal. Of 8 X-linked genes identified previously in L. cuprina, 6 were orthologs of Drosophila 4 th -chromosome genes [2]. The X-linked genes were expressed equally in males and females. Here we identify an additional 51 X-linked genes and show that most are dosage compensated. Orthologs of 49 of the 59 X-linked genes are on the 4 th  chromosome in D. melanogaster. Because painting of fourth (Pof) is important for expression of Drosophila 4 th -chromosome genes [3], we used Cas9 to make a loss-of-function knockin mutation in an L. cuprina Pof ortholog we call no blokes (nbl). Homozygous nbl males derived from homozygous nbl mothers die at the late pupal stage. Homozygous nbl females are viable, fertile, and live longer than heterozygous nbl females. RNA expression of most X-linked genes was reduced in homozygous nbl male pupae and to a lesser extent in nbl females compared to heterozygous siblings. The results suggest that NBL could be important for X chromosome dosage compensation in L. cuprina. NBL may also facilitate gene expression in the heterochromatic environment of the X chromosome in both sexes. This study supports the hypothesis on the origin of the Drosophila 4 th chromosome and that a POF-like protein was required for normal gene expression on the ancient X chromosome.}, number={12}, journal={Current Biology}, publisher={Elsevier BV}, author={Davis, Rebecca J. and Belikoff, Esther J. and Scholl, Elizabeth H. and Li, Fang and Scott, Maxwell J.}, year={2018}, month={Jun}, pages={1987–1992.e3} } @article{davis_belikoff_scholl_li_scott_2018, title={no blokes Is Essential for Male Viability and X Chromosome Gene Expression in the Australian Sheep Blowfly}, volume={28}, url={http://www.sciencedirect.com/science/article/pii/S0960982218306079}, DOI={https://doi.org/10.1016/j.cub.2018.05.005}, abstractNote={It has been hypothesized that the Drosophila 4th chromosome is derived from an ancient X chromosome [1]. In the Australian sheep blowfly, Lucilia cuprina, the heterochromatic X chromosome contains few active genes and orthologs of Drosophila X-linked genes are autosomal. Of 8 X-linked genes identified previously in L. cuprina, 6 were orthologs of Drosophila 4th-chromosome genes [2]. The X-linked genes were expressed equally in males and females. Here we identify an additional 51 X-linked genes and show that most are dosage compensated. Orthologs of 49 of the 59 X-linked genes are on the 4th chromosome in D. melanogaster. Because painting of fourth (Pof) is important for expression of Drosophila 4th-chromosome genes [3], we used Cas9 to make a loss-of-function knockin mutation in an L. cuprina Pof ortholog we call no blokes (nbl). Homozygous nbl males derived from homozygous nbl mothers die at the late pupal stage. Homozygous nbl females are viable, fertile, and live longer than heterozygous nbl females. RNA expression of most X-linked genes was reduced in homozygous nbl male pupae and to a lesser extent in nbl females compared to heterozygous siblings. The results suggest that NBL could be important for X chromosome dosage compensation in L. cuprina. NBL may also facilitate gene expression in the heterochromatic environment of the X chromosome in both sexes. This study supports the hypothesis on the origin of the Drosophila 4th chromosome and that a POF-like protein was required for normal gene expression on the ancient X chromosome.}, number={12}, journal={Current Biology}, author={Davis, Rebecca J. and Belikoff, Esther J. and Scholl, Elizabeth H. and Li, Fang and Scott, Maxwell J.}, year={2018}, pages={1987–1992.e3} } @article{scott_gould_lorenzen_grubbs_edwards_o’brochta_2017, title={Agricultural production: assessment of the potential use of Cas9-mediated gene drive systems for agricultural pest control}, volume={5}, ISSN={2329-9460 2329-9037}, url={http://dx.doi.org/10.1080/23299460.2017.1410343}, DOI={10.1080/23299460.2017.1410343}, abstractNote={ABSTRACTTo highlight how gene drives could be useful for control of agricultural insect pests, we selected species that are pests of animals (New World screwworm), plants (spotted wing Drosophila, ...}, number={sup1}, journal={Journal of Responsible Innovation}, publisher={Informa UK Limited}, author={Scott, Maxwell J. and Gould, Fred and Lorenzen, Marcé and Grubbs, Nathaniel and Edwards, Owain and O’Brochta, David}, year={2017}, month={Dec}, pages={S98–S120} } @article{yan_linger_scott_2017, title={Building early-larval sexing systems for genetic control of the Australian sheep blow fly Lucilia cuprina using two constitutive promoters}, volume={7}, ISSN={2045-2322}, url={http://dx.doi.org/10.1038/s41598-017-02763-4}, DOI={10.1038/s41598-017-02763-4}, abstractNote={AbstractTransgenic sexing strains (TSS) that carry conditional female lethal genes are advantageous for genetic control programs based on the sterile insect technique (SIT). It is desirable if females die early in development as larval diet is a major cost for mass production facilities. This can be achieved by using a gene promoter that is only active in embryos to drive expression of the tetracycline transactivator (tTA), the transcription factor commonly used in two-component TSS. While an embryo-specific promoter is ideal it may not be essential for assembling an effective TSS as tTA can be repressed by addition of tetracycline to the diet at larval and/or adult stages. Here we have investigated this idea by isolating and employing the promoters from the Lucilia spitting image and actin 5C genes to drive tTA expression in embryos and later stages. L. cuprina TSS with the tTA drivers and tTA-regulated tetO-Lshid effectors produced only females when raised on a limited tetracycline diet. The Lshid transgene contains a sex-specific intron and as a consequence only females produce LsHID protein. TSS females died at early larval stages, which makes the lines advantageous for an SIT program.}, number={1}, journal={Scientific Reports}, publisher={Springer Science and Business Media LLC}, author={Yan, Ying and Linger, Rebecca J. and Scott, Maxwell J.}, year={2017}, month={May} } @article{scott_concha_welch_phillips_skoda_2017, title={Review of research advances in the screwworm eradication program over the past 25 years}, volume={164}, ISSN={0013-8703 1570-7458}, url={http://dx.doi.org/10.1111/eea.12607}, DOI={10.1111/eea.12607}, abstractNote={AbstractNew World screwworms, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae: Chrysomyinae), are devastating pests of warm‐blooded animals that cause significant economic damage to livestock. The successful campaign to eradicate screwworms from continental North America, led by the US Department of Agriculture and using the sterile insect technique, continues to receive research support that has resulted in improved technologies for all aspects of the program. The process and ingredients for mass‐rearing screwworms is more efficient and sustainable, and there is now a standardized protocol for developing new strains used in mass rearing. Cryopreservation of screwworm embryos allows strains to be preserved and recovered if necessary and also reduces rearing requirements for backup and research strains. Sterile fly release procedures and equipment have been updated leading to optimized sterile fly release rates. Surveillance for screwworm infestations and outbreaks have incorporated new trap designs, habitat analysis, and molecular genetic techniques that enhance monitoring the progress of the program as well as early detection and response to outbreaks. Genetic analyses of screwworm populations across their current range have increased the understanding of genetic differentiation, which may aide in developing new strains and determining the geographic origin of screwworms causing outbreaks when they occur. The ability to release only sterile males, which has been a goal of the program for over 60 years, has recently been accomplished through the development of transgenic sexing strains. The strains carry a conditional female lethal gene and are comparable to the wild‐type strain for several biological parameters that are important for mass production and performance in the field. The strains should improve efficiency of population suppression of the current and future eradication and prevention programs against screwworms. These research advances as well as future considerations are presented.}, number={3}, journal={Entomologia Experimentalis et Applicata}, publisher={Wiley}, author={Scott, Maxwell J. and Concha, Carolina and Welch, John B. and Phillips, Pamela L. and Skoda, Steven R.}, year={2017}, month={Aug}, pages={226–236} } @article{dearden_gemmell_mercier_lester_scott_newcomb_buckley_jacobs_goldson_penman_2017, title={The potential for the use of gene drives for pest control in New Zealand: a perspective}, volume={48}, ISSN={0303-6758 1175-8899}, url={http://dx.doi.org/10.1080/03036758.2017.1385030}, DOI={10.1080/03036758.2017.1385030}, abstractNote={ABSTRACT Genetic technologies such as gene editing and gene drive systems have recently emerged as potential tools for pest control. Gene drives, in particular, have been described as potential solutions to the pest problems that beset New Zealand. Here we describe the current state of gene drive technologies and present a series of examples to examine the potential benefits and problems arising from gene drive approaches for pest control in New Zealand. We consider the risks and barriers, both biological and social, that would need to be addressed to deploy such systems against our key pests with particular reference to the unique characteristics of New Zealand’s biota, environment and peoples. Gene drives are a potentially useful technology for the eradication of pests in New Zealand but a great deal of research and understanding, as well as public acceptance, is required before they can be implemented.}, number={4}, journal={Journal of the Royal Society of New Zealand}, publisher={Informa UK Limited}, author={Dearden, Peter K. and Gemmell, Neil J. and Mercier, Ocean R. and Lester, Philip J. and Scott, Maxwell J. and Newcomb, Richard D. and Buckley, Thomas R. and Jacobs, Jeanne M. E. and Goldson, Stephen G. and Penman, David R.}, year={2017}, month={Oct}, pages={225–244} } @article{anstead_batterham_korhonen_young_hall_bowles_richards_scott_gasser_2016, title={A blow to the fly — Lucilia cuprina draft genome and transcriptome to support advances in biology and biotechnology}, volume={34}, ISSN={0734-9750}, url={http://dx.doi.org/10.1016/j.biotechadv.2016.02.009}, DOI={10.1016/j.biotechadv.2016.02.009}, abstractNote={The blow fly, Lucilia cuprina (Wiedemann, 1830) is a parasitic insect of major global economic importance. Maggots of this fly parasitize the skin of animal hosts, feed on excretions and tissues, and cause severe disease (flystrike or myiasis). Although there has been considerable research on L. cuprina over the years, little is understood about the molecular biology, biochemistry and genetics of this parasitic fly, as well as its relationship with its hosts and the disease that it causes. This situation might change with the recent report of the draft genome and transcriptome of this blow fly, which has given new and global insights into its biology, interactions with the host animal and aspects of insecticide resistance at the molecular level. This genomic resource will likely enable many fundamental and applied research areas in the future. The present article gives a background on L. cuprina and myiasis, a brief account of past and current treatment, prevention and control approaches, and provides a perspective on the impact that the L. cuprina genome should have on future research of this and related parasitic flies, and the design of new and improved interventions for myiasis.}, number={5}, journal={Biotechnology Advances}, publisher={Elsevier BV}, author={Anstead, Clare A. and Batterham, Philip and Korhonen, Pasi K. and Young, Neil D. and Hall, Ross S. and Bowles, Vernon M. and Richards, Stephen and Scott, Maxwell J. and Gasser, Robin B.}, year={2016}, month={Sep}, pages={605–620} } @article{concha_palavesam_guerrero_sagel_li_osborne_hernandez_pardo_quintero_vasquez_et al._2016, title={A transgenic male-only strain of the New World screwworm for an improved control program using the sterile insect technique}, volume={14}, ISSN={1741-7007}, url={http://dx.doi.org/10.1186/s12915-016-0296-8}, DOI={10.1186/s12915-016-0296-8}, abstractNote={The New World screwworm, Cochliomyia hominivorax, is a devastating pest of livestock endemic to subtropical and tropical regions of the Western hemisphere. The larvae of this species feed on the tissue of living animals, including man, and can cause death if untreated. Over 60 years ago, the sterile insect technique (SIT) was developed with the aim of eradicating this pest, initially from Florida but subsequently from all of North and Central America. From the outset it was appreciated that SIT would be more efficient if only sterile males were released in the field, but this was not possible until now. Here, we report on the development and evaluation of the first sexing strains of C. hominivorax that produce only males when raised on diet without tetracycline. Transgenic lines have been developed that possess a tetracycline repressible female-lethal genetic system. Ten of these lines show high female lethality at the late larval/pupal stages and three of them present dominant female lethality. Most of the lines were comparable to the wild type parental strain in several fitness parameters that are relevant to mass rearing in a production facility. Further, three lines performed well in male mating success and male competition assays, suggesting they would be sexually competitive in the field. Consequently, one transgenic line has been selected by the New World Screwworm Program for evaluation under mass rearing conditions. We conclude that the promising characteristics of the selected sexing strains may contribute to reduce production costs for the existing eradication program and provide more efficient population suppression, which should make a genetic control program more economical in regions were C. hominivorax remains endemic.}, number={1}, journal={BMC Biology}, publisher={Springer Science and Business Media LLC}, author={Concha, Carolina and Palavesam, Azhahianambi and Guerrero, Felix D. and Sagel, Agustin and Li, Fang and Osborne, Jason A. and Hernandez, Yillian and Pardo, Trinidad and Quintero, Gladys and Vasquez, Mario and et al.}, year={2016}, month={Aug} } @article{concha_palavesam_guerrero_sagel_li_osborne_hernandez_pardo_quintero_vasquez_et al._2016, title={A transgenic male-only strain of the New World screwworm for an improved control program using the sterile insect technique}, volume={14}, number={1}, journal={BMC biology}, publisher={BioMed Central}, author={Concha, Carolina and Palavesam, Azhahianambi and Guerrero, Felix D and Sagel, Agustin and Li, Fang and Osborne, Jason A and Hernandez, Yillian and Pardo, Trinidad and Quintero, Gladys and Vasquez, Mario and et al.}, year={2016}, pages={72} } @article{scott_benedict_2016, title={Concept and history of genetic control}, journal={Genetic Control of Malaria and Dengue}, author={Scott, M. J. and Benedict, M. Q.}, year={2016}, pages={31–54} } @article{schwartz_truglio_scott_fitzsimons_2016, title={Long-Term Memory inDrosophilaIs Influenced by Histone Deacetylase HDAC4 Interacting with SUMO-Conjugating Enzyme Ubc9}, volume={203}, ISSN={1943-2631}, url={http://dx.doi.org/10.1534/genetics.115.183194}, DOI={10.1534/genetics.115.183194}, abstractNote={AbstractHDAC4 is a potent memory repressor with overexpression of wild type or a nuclear-restricted mutant resulting in memory deficits. Interestingly, reduction of HDAC4 also impairs memory via an as yet unknown mechanism. Although histone deacetylase family members are important mediators of epigenetic mechanisms in neurons, HDAC4 is predominantly cytoplasmic in the brain and there is increasing evidence for interactions with nonhistone proteins, suggesting HDAC4 has roles beyond transcriptional regulation. To that end, we performed a genetic interaction screen in Drosophila and identified 26 genes that interacted with HDAC4, including Ubc9, the sole SUMO E2-conjugating enzyme. RNA interference-induced reduction of Ubc9 in the adult brain impaired long-term memory in the courtship suppression assay, a Drosophila model of associative memory. We also demonstrate that HDAC4 and Ubc9 interact genetically during memory formation, opening new avenues for investigating the mechanisms through which HDAC4 regulates memory formation and other neurological processes.}, number={3}, journal={Genetics}, publisher={Oxford University Press (OUP)}, author={Schwartz, Silvia and Truglio, Mauro and Scott, Maxwell J and Fitzsimons, Helen L}, year={2016}, month={Jul}, pages={1249–1264} } @article{linger_belikoff_yan_li_wantuch_fitzsimons_scott_2016, title={Towards next generation maggot debridement therapy: transgenic Lucilia sericata larvae that produce and secrete a human growth factor}, volume={16}, ISSN={1472-6750}, url={http://dx.doi.org/10.1186/s12896-016-0263-z}, DOI={10.1186/s12896-016-0263-z}, abstractNote={Diabetes and its concurrent complications impact a significant proportion of the population of the US and create a large financial burden on the American health care system. FDA-approved maggot debridement therapy (MDT), the application of sterile laboratory-reared Lucilia sericata (green bottle fly) larvae to wounds, is a cost-effective and successful treatment for diabetic foot ulcers and other medical conditions. Human platelet derived growth factor-BB (PDGF-BB) is a secreted dimeric peptide growth factor that binds the PDGF receptor. PDGF-BB stimulates cell proliferation and survival, promotes wound healing, and has been investigated as a possible topical treatment for non-healing wounds. Genetic engineering has allowed for expression and secretion of human growth factors and other proteins in transgenic insects. Here, we present a novel concept in MDT technology that combines the established benefits of MDT with the power of genetic engineering to promote healing. The focus of this study is to create and characterize strains of transgenic L. sericata that express and secrete PDGF-BB at detectable levels in adult hemolymph, whole larval lysate, and maggot excretions/ secretions (ES), with potential for clinical utility in wound healing. We have engineered and confirmed transgene insertion in several strains of L. sericata that express human PDGF-BB. Using a heat-inducible promoter to control the pdgf-b gene, pdgf-b mRNA was detected via semi-quantitative PCR upon heat shock. PDGF-BB protein was also detectable in larval lysates and adult hemolymph but not larval ES. An alternative, tetracycline-repressible pdgf-b system mediated expression of pdgf-b mRNA when maggots were raised on diet that lacked tetracycline. Further, PDGF-BB protein was readily detected in whole larval lysate as well as larval ES. Here we show robust, inducible expression and production of human PDGF-BB protein from two conditional expression systems in transgenic L. sericata larvae. The tetracycline-repressible system appears to be the most promising as PDGF-BB protein was detectable in larval ES following induction. Our system could potentially be used to deliver a variety of growth factors and anti-microbial peptides to the wound environment with the aim of enhancing wound healing, thereby improving patient outcome in a cost-effective manner.}, number={1}, journal={BMC Biotechnology}, publisher={Springer Science and Business Media LLC}, author={Linger, Rebecca J. and Belikoff, Esther J. and Yan, Ying and Li, Fang and Wantuch, Holly A. and Fitzsimons, Helen L. and Scott, Maxwell J.}, year={2016}, month={Mar} } @article{yan_scott_2015, title={A transgenic embryonic sexing system for the Australian sheep blow fly Lucilia cuprina}, volume={5}, ISSN={2045-2322}, url={http://dx.doi.org/10.1038/srep16090}, DOI={10.1038/srep16090}, abstractNote={AbstractGenetic approaches, including the sterile insect technique (SIT), have previously been considered for control of the Australian sheep blow fly Lucilia cuprina, a major pest of sheep. In an SIT program, females consume 50% of the diet but are ineffective as control agents and compete with females in the field for mating with sterile males, thereby decreasing the efficiency of the program. Consequently, transgenic sexing strains of L. cuprina were developed that produce 100% males when raised on diet that lacks tetracycline. However, as females die mostly at the pupal stage, rearing costs would not be significantly reduced. Here we report the development of transgenic embryonic sexing strains of L. cuprina. In these strains, the Lsbnk cellularization gene promoter drives high levels of expression of the tetracycline transactivator (tTA) in the early embryo. In the absence of tetracycline, tTA activates expression of the Lshid proapoptotic gene, leading to death of the embryo. Sex-specific RNA splicing of Lshid transcripts ensures that only female embryos die. Embryonic sexing strains were also made by combining the Lsbnk-tTA and tetO-Lshid components into a single gene construct, which will facilitate transfer of the technology to other major calliphorid livestock pests.}, number={1}, journal={Scientific Reports}, publisher={Springer Science and Business Media LLC}, author={Yan, Ying and Scott, Maxwell J.}, year={2015}, month={Nov} } @article{li_scott_2016, title={CRISPR/Cas9-mediated mutagenesis of the white and Sex lethal loci in the invasive pest, Drosophila suzukii}, volume={469}, ISSN={0006-291X}, url={http://dx.doi.org/10.1016/j.bbrc.2015.12.081}, DOI={10.1016/j.bbrc.2015.12.081}, abstractNote={Drosophila suzukii (commonly called spotted wing Drosophila) is an invasive pest of soft-skinned fruit (e.g. blueberries, strawberries). A high quality reference genome sequence is available but functional genomic tools, such as used in Drosophila melanogaster, remain to be developed. In this study we have used the CRISPR/Cas9 system to introduce site-specific mutations in the D. suzukii white (w) and Sex lethal (Sxl) genes. Hemizygous males with w mutations develop white eyes and the mutant genes are transmissible to the next generation. Somatic mosaic females that carry mutations in the Sxl gene develop abnormal genitalia and reproductive tissue. The D. suzukii Sxl gene could be an excellent target for a Cas9-mediated gene drive to suppress populations of this highly destructive pest.}, number={4}, journal={Biochemical and Biophysical Research Communications}, publisher={Elsevier BV}, author={Li, Fang and Scott, Maxwell J.}, year={2016}, month={Jan}, pages={911–916} } @misc{scott_benedict_2016, title={Concept and History of Genetic Control}, url={http://dx.doi.org/10.1016/B978-0-12-800246-9.00002-8}, DOI={10.1016/B978-0-12-800246-9.00002-8}, abstractNote={Abstract Genetic control of insects is an established method, mainly for insects that are important crop and veterinary pests such as medflies and screwworm. Efforts to use the same technologies against insects of medical importance, especially mosquitoes, have had limited success. The successes against mosquitoes have been accomplished using forms of both conventional and modern methods, both of which are promising. In this chapter, we provide highlights of the development of genetic control of agricultural pests and describe how the development of methods against mosquitoes reflects those advances. While admiring successful genetic control programs is motivating, we suggest that much can also be learned from both past successful and failed efforts, as doing so will increase our ability to improve future activities.}, journal={Genetic Control of Malaria and Dengue}, publisher={Elsevier}, author={Scott, Maxwell J. and Benedict, Mark Q.}, year={2016}, pages={31–54} } @inbook{scott_benedict_2015, title={Concept and history of genetic control}, booktitle={Genetic control of malaria and dengue}, publisher={Elsevier}, author={Scott, Maxwell J. and Benedict, Mark Q.}, year={2015}, pages={31–54} } @article{linger_belikoff_scott_2015, title={Dosage Compensation of X-Linked Muller Element F Genes but Not X-Linked Transgenes in the Australian Sheep Blowfly}, volume={10}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0141544}, DOI={10.1371/journal.pone.0141544}, abstractNote={In most animals that have X and Y sex chromosomes, chromosome-wide mechanisms are used to balance X-linked gene expression in males and females. In the fly Drosophila melanogaster, the dosage compensation mechanism also generally extends to X-linked transgenes. Over 70 transgenic lines of the Australian sheep blowfly Lucilia cuprina have been made as part of an effort to develop male-only strains for a genetic control program of this major pest of sheep. All lines carry a constitutively expressed fluorescent protein marker gene. In all 12 X-linked lines, female larvae show brighter fluorescence than male larvae, suggesting the marker gene is not dosage compensated. This has been confirmed by quantitative RT-PCR for selected lines. To determine if endogenous X-linked genes are dosage compensated, we isolated 8 genes that are orthologs of genes that are on the fourth chromosome in D. melanogaster. Recent evidence suggests that the D. melanogaster fourth chromosome, or Muller element F, is the ancestral X chromosome in Diptera that has reverted to an autosome in Drosophila species. We show by quantitative PCR of male and female DNA that 6 of the 8 linkage group F genes reside on the X chromosome in L. cuprina. The other two Muller element F genes were found to be autosomal in L. cuprina, whereas two Muller element B genes were found on the same region of the X chromosome as the L. cuprina orthologs of the D. melanogaster Ephrin and gawky genes. We find that the L. cuprina X chromosome genes are equally expressed in males and females (i.e., fully dosage compensated). Thus, unlike in Drosophila, it appears that the Lucilia dosage compensation system is specific for genes endogenous to the X chromosome and cannot be co-opted by recently arrived transgenes.}, number={10}, journal={PLOS ONE}, publisher={Public Library of Science (PLoS)}, author={Linger, Rebecca J. and Belikoff, Esther J. and Scott, Maxwell J.}, editor={Marais, Gabriel ABEditor}, year={2015}, month={Oct}, pages={e0141544} } @article{sandeman_levot_heath_james_greeff_scott_batterham_bowles_2014, title={Control of the sheep blowfly in Australia and New Zealand – are we there yet?}, volume={44}, ISSN={0020-7519}, url={http://dx.doi.org/10.1016/j.ijpara.2014.08.009}, DOI={10.1016/j.ijpara.2014.08.009}, abstractNote={The last 50 years of research into infections in Australia and New Zealand caused by larvae of the sheep blowfly, Lucilia cuprina, have significantly advanced our understanding of this blowfly and its primary host, the sheep. However, apart from some highly effective drugs it could be argued that no new control methodologies have resulted. This review addresses the major areas of sheep blowfly research over this period describing the significant outcomes and analyses, and what is still required to produce new commercial control technologies. The use of drugs against this fly species has been very successful but resistance has developed to almost all current compounds. Integrated pest management is becoming basic to control, especially in the absence of mulesing, and has clearly benefited from computer-aided technologies. Biological control has more challenges but natural and perhaps transformed biopesticides offer possibilities for the future. Experimental vaccines have been developed but require further analysis of antigens and formulations to boost protection. Genetic technologies may provide potential for long-term control through more rapid indirect selection of sheep less prone to flystrike. Finally in the future, genetic analysis of the fly may allow suppression and perhaps eradication of blowfly populations or identification of new and more viable targets for drug and vaccine intervention. Clearly all these areas of research offer potential new controls but commercial development is perhaps inhibited by the success of current chemical insecticides and certainly requires a significant additional injection of resources.}, number={12}, journal={International Journal for Parasitology}, publisher={Elsevier BV}, author={Sandeman, R.M. and Levot, G.W. and Heath, A.C.G. and James, P.J. and Greeff, J.C. and Scott, M.J. and Batterham, P. and Bowles, V.M.}, year={2014}, month={Oct}, pages={879–891} } @article{sandeman_levot_heath_james_greeff_scott_batterham_bowles_2014, title={Control of the sheep blowfly in Australia and New Zealand--are we there yet?}, volume={44}, number={12}, journal={International journal for parasitology}, publisher={Pergamon}, author={Sandeman, RM and Levot, GW and Heath, ACG and James, PJ and Greeff, JC and Scott, MJ and Batterham, P and Bowles, VM}, year={2014}, pages={879–891} } @article{scott_2014, title={Development and evaluation of male-only strains of the Australian sheep blowfly, Lucilia cuprina}, volume={15}, ISSN={2730-6844}, url={http://dx.doi.org/10.1186/1471-2156-15-S2-S3}, DOI={10.1186/1471-2156-15-s2-s3}, abstractNote={AbstractThe Australian sheep blowflyLucilia cuprina(Wiedemann) is a major pest of sheep in Australia and New Zealand. From the 1960s to the 1980s there was a major effort to develop "field female killing" or FFK strains ofL. cuprinathat could be used for a cost-effective genetic control program. The FFK strains carried eye color mutations that were lethal to females in the field but not under conditions in the mass rearing facility. Males did not die in the field as normal copies of the eye color genes had been translocated to the Y chromosome and an autosome. Although the FFK strains showed some promise in field tests, a genetic control program in mainland Australia was never implemented for several reasons including instability of the FFK strains during mass rearing. A stable transgenic strain ofL. cuprinathat carried one or more dominant repressible female lethal genes offered the potential for efficient genetic control of blowfly populations. Here I review our research on tetracycline-repressible female lethal genetic systems,Luciliagerm-line transformation and sex determination genes that ultimately led to the successful development of transgenic "male-only" strains ofL. cuprina. The technology developed forL. cuprinashould be directly transferable to other blowfly livestock pests includingL. sericataand the New World and Old World screwworm. 29}, number={S2}, journal={BMC Genomic Data}, publisher={Springer Science and Business Media LLC}, author={Scott, Maxwell J}, year={2014}, month={Dec} } @article{edman_linger_belikoff_li_sze_tarone_scott_2014, title={Functional characterization of calliphorid cell death genes and cellularization gene promoters for controlling gene expression and cell viability in early embryos}, volume={24}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1111/imb.12135}, DOI={10.1111/imb.12135}, abstractNote={AbstractThe New World screwworm fly, Cochliomyia hominivorax, and the Australian sheep blow fly, Lucilia cuprina, are major pests of livestock. The sterile insect technique was used to eradicate C. hominivorax from North and Central America. This involved area‐wide releases of male and female flies that had been sterilized by radiation. Genetic systems have been developed for making ‘male‐only’ strains that would improve the efficiency of genetic control of insect pests. One system involves induction of female lethality in embryos through activation of a pro‐apoptotic gene by the tetracycline‐dependent transactivator. Sex‐specific expression is achieved using an intron from the transformer gene, which we previously isolated from several calliphorids. In the present study, we report the isolation of the promoters from the C. hominivorax slam and Lucilia sericata bnk cellularization genes and show that these promoters can drive expression of a GFP reporter gene in early embryos of transgenic L. cuprina. Additionally, we report the isolation of the L. sericata pro‐apoptotic hid and rpr genes, identify conserved motifs in the encoded proteins and determine the relative expression of these genes at different stages of development. We show that widespread expression of the L. sericata pro‐apoptotic genes was lethal in Drosophila melanogaster. The isolated gene promoters and pro‐apoptotic genes could potentially be used to build transgenic embryonic sexing strains of calliphorid livestock pests.}, number={1}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Edman, R. M. and Linger, R. J. and Belikoff, E. J. and Li, F. and Sze, S.‐H. and Tarone, A. M. and Scott, M. J.}, year={2014}, month={Sep}, pages={58–70} } @article{attardo_abila_auma_baumann_benoit_brelsfoard_ribeiro_cotton_pham_darby_et al._2014, title={Genome Sequence of the Tsetse Fly ( Glossina morsitans ): Vector of African Trypanosomiasis}, volume={344}, ISSN={0036-8075 1095-9203}, url={http://dx.doi.org/10.1126/science.1249656}, DOI={10.1126/science.1249656}, abstractNote={Africa's Bane Tsetse are blood-feeding, fast-flying flies that transmit a range of Trypanosoma spp. protozoan pathogens, which cause sleeping sickness in humans and their nagana in their livestock. The International Glossina Genome Initiative (p. 380 ) sequenced the genome of Glossina morsitans and identified the genes for many attributes of the tsetse's remarkable biology, including viviparity and the expression of analogs of mammalian milk proteins. Tsetse are host to several specific symbionts that appear to synthesize essential nutrients for the fly and also to hitherto undiscovered parasitoid-derived viruses. Deeper exploration of this genome will reveal what makes these fly species so host- and trypanosome specific. }, number={6182}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Attardo, Geoffrey M. and Abila, Patrick P. and Auma, Joanna E. and Baumann, Aaron A. and Benoit, Joshua B. and Brelsfoard, Corey L. and Ribeiro, José M. C. and Cotton, James A. and Pham, Daphne Q. D. and Darby, Alistair C. and et al.}, year={2014}, month={Apr}, pages={380–386} } @article{watanabe_hattori_berriman_lehane_hall_solano_aksoy_hide_toure_attardo_et al._2014, title={Genome sequence of the tsetse fly (Glossina morsitans): vector of African trypanosomiasis}, volume={344}, number={6182}, journal={Science}, author={Watanabe, J. and Hattori, M. and Berriman, M. and Lehane, M. J. and Hall, N. and Solano, P. and Aksoy, S. and Hide, W. and Toure, Y. and Attardo, G. M. and et al.}, year={2014}, pages={380–386} } @article{attardo_abila_auma_baumann_benoit_brelsfoard_ribeiro_cotton_pham_darby_et al._2014, title={Genome sequence of the tsetse fly (Glossina morsitans): vector of African trypanosomiasis}, volume={344}, number={6182}, journal={Science}, publisher={American Association for the Advancement of Science}, author={Attardo, Geoffrey M and Abila, Patrick P and Auma, Joanna E and Baumann, Aaron A and Benoit, Joshua B and Brelsfoard, Corey L and Ribeiro, José MC and Cotton, James A and Pham, Daphne QD and Darby, Alistair C and et al.}, year={2014}, pages={380–386} } @article{scott_morrison_simmons_2014, title={Transgenic approaches for sterile insect control of dipteran livestock pests and lepidopteran crop pests}, journal={Transgenic insects: techniques and applications}, author={Scott, M. J. and Morrison, N. I. and Simmons, G. S.}, year={2014}, pages={152–167} } @misc{scott_morrison_simmons_2014, title={Transgenic approaches for sterile insect control of dipteran livestock pests and lepidopteran crop pests.}, ISBN={9781780644516}, url={http://dx.doi.org/10.1079/9781780644516.0152}, DOI={10.1079/9781780644516.0152}, abstractNote={AbstractThis chapter focuses on the various transgenic approaches in the control of dipteran livestock pests and lepidopteran crop pests based on the sterile insect technique.}, journal={Transgenic insects: techniques and applications}, publisher={CABI}, author={Scott, M. J. and Morrison, N. I. and Simmons, G. S.}, year={2014}, pages={152–167} } @inbook{scott_morrison_simmons_benedict_others_2014, title={Transgenic approaches for sterile insect control of dipteran livestock pests and lepidopteran crop pests.}, booktitle={Transgenic Insects: Techniques and Applications}, publisher={CABI}, author={Scott, Maxwell J and Morrison, Neil I and Simmons, Gregory S and Benedict, MQ and others}, year={2014}, pages={152} } @article{li_wantuch_linger_belikoff_scott_2014, title={Transgenic sexing system for genetic control of the Australian sheep blow fly Lucilia cuprina}, volume={51}, ISSN={0965-1748}, url={http://dx.doi.org/10.1016/j.ibmb.2014.06.001}, DOI={10.1016/j.ibmb.2014.06.001}, abstractNote={The New World screwworm and the Australian sheep blowfly Lucilia cuprina are devastating pests of livestock. The larvae of these species feed on the tissue of the living animal and can cause death if untreated. The sterile insect technique or SIT was used to eradicate screwworm from North and Central America. This inspired efforts to develop strains containing complex chromosomal rearrangements for genetic control of L. cuprina in Australia. Although one field trial was promising, the approach was abandoned due to costs and difficulties in mass rearing the strain. As the efficiency of SIT can be significantly increased if only sterile males are released, we have developed transgenic strains of L. cuprina that carry a dominant tetracycline repressible female lethal genetic system. Lethality is due to overexpression of an auto-regulated tetracycline repressible transactivator (tTA) gene and occurs mostly at the pupal stage. Dominant female lethality was achieved by replacing the Drosophila hsp70 core promoter with a Lucilia hsp70 core promoter-5'UTR for tTA overexpression. The strains carry a dominant strongly expressed marker that will facilitate identification in the field. Interestingly, the sexes could be reliably sorted by fluorescence or color from the early first instar larval stage as females that overexpress tTA also overexpress the linked marker gene. Male-only strains of L. cuprina developed in this study could form the basis for a future genetic control program. Moreover, the system developed for L. cuprina should be readily transferrable to other major calliphorid livestock pests including the New and Old World screwworm.}, journal={Insect Biochemistry and Molecular Biology}, publisher={Elsevier BV}, author={Li, Fang and Wantuch, Holly A. and Linger, Rebecca J. and Belikoff, Esther J. and Scott, Maxwell J.}, year={2014}, month={Aug}, pages={80–88} } @article{li_vensko_belikoff_scott_2013, title={Conservation and Sex-Specific Splicing of the transformer Gene in the Calliphorids Cochliomyia hominivorax, Cochliomyia macellaria and Lucilia sericata}, volume={8}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0056303}, DOI={10.1371/journal.pone.0056303}, abstractNote={Transformer (TRA) promotes female development in several dipteran species including the Australian sheep blowfly Lucilia cuprina, the Mediterranean fruit fly, housefly and Drosophila melanogaster. tra transcripts are sex-specifically spliced such that only the female form encodes full length functional protein. The presence of six predicted TRA/TRA2 binding sites in the sex-specific female intron of the L. cuprina gene suggested that tra splicing is auto-regulated as in medfly and housefly. With the aim of identifying conserved motifs that may play a role in tra sex-specific splicing, here we have isolated and characterized the tra gene from three additional blowfly species, L. sericata, Cochliomyia hominivorax and C. macellaria. The blowfly adult male and female transcripts differ in the choice of splice donor site in the first intron, with males using a site downstream of the site used in females. The tra genes all contain a single TRA/TRA2 site in the male exon and a cluster of four to five sites in the male intron. However, overall the sex-specific intron sequences are poorly conserved in closely related blowflies. The most conserved regions are around the exon/intron junctions, the 3′ end of the intron and near the cluster of TRA/TRA2 sites. We propose a model for sex specific regulation of tra splicing that incorporates the conserved features identified in this study. In L. sericata embryos, the male tra transcript was first detected at around the time of cellular blastoderm formation. RNAi experiments showed that tra is required for female development in L. sericata and C. macellaria. The isolation of the tra gene from the New World screwworm fly C. hominivorax, a major livestock pest, will facilitate the development of a “male-only” strain for genetic control programs.}, number={2}, journal={PLoS ONE}, publisher={Public Library of Science (PLoS)}, author={Li, Fang and Vensko, Steven P. and Belikoff, Esther J. and Scott, Maxwell J.}, editor={Tear, GuyEditor}, year={2013}, month={Feb}, pages={e56303} } @article{scott_pimsler_tarone_2014, title={Sex Determination Mechanisms in the Calliphoridae (Blow Flies)}, volume={8}, ISSN={1661-5425 1661-5433}, url={http://dx.doi.org/10.1159/000357132}, DOI={10.1159/000357132}, abstractNote={The Calliphoridae or blow flies are a family of insects that occupy diverse habitats and perform important ecological roles, particularly the decomposition of animal remains. Some Calliphoridae species are also important in the forensic sciences, in agriculture (e.g. as livestock pests) and in medicine (e.g. maggot therapy). Calliphoridae provide striking examples in support of the hypothesis that sex determination regulatory gene hierarchies evolve in the reverse order, with the gene at the top being the most recently added. Unlike the model fly Drosophila melanogaster, where sex is determined by the number of X chromosomes, in the Australian sheep blow fly (Lucilia cuprina) sex is determined by a Y-linked male-determining gene (M). A different regulatory system appears to operate in the hairy maggot blow fly (Chrysomya rufifacies) where the maternal genotype determines sex. It is hypothesized that females heterozygous for a dominant female-determining factor (F/f) produce only female offspring and homozygous f/f females produce only sons. The bottom of the regulatory hierarchy appears to be the same in D. melanogaster and L. cuprina, with sex-specific splicing of doublesex transcripts being controlled by the female-specific Transformer (TRA) protein. We discuss a model that has been proposed for how tra transcripts are sex-specifically spliced in calliphorids, which is very different from D. melanogaster.}, number={1-3}, journal={Sexual Development}, publisher={S. Karger AG}, author={Scott, M.J. and Pimsler, M.L. and Tarone, A.M.}, year={2014}, pages={29–37} } @article{scott_pimsler_tarone_2013, title={Sex determination mechanisms in the Calliphoridae (blow flies)}, volume={8}, number={1-3}, journal={Sexual Development}, publisher={Karger Publishers}, author={Scott, MJ and Pimsler, ML and Tarone, AM}, year={2013}, pages={29–37} } @article{fitzsimons_schwartz_given_scott_2013, title={The Histone Deacetylase HDAC4 Regulates Long-Term Memory in Drosophila}, volume={8}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0083903}, DOI={10.1371/journal.pone.0083903}, abstractNote={A growing body of research indicates that pharmacological inhibition of histone deacetylases (HDACs) correlates with enhancement of long-term memory and current research is concentrated on determining the roles that individual HDACs play in cognitive function. Here, we investigate the role of HDAC4 in long-term memory formation in Drosophila. We show that overexpression of HDAC4 in the adult mushroom body, an important structure for memory formation, resulted in a specific impairment in long-term courtship memory, but had no affect on short-term memory. Overexpression of an HDAC4 catalytic mutant also abolished LTM, suggesting a mode of action independent of catalytic activity. We found that overexpression of HDAC4 resulted in a redistribution of the transcription factor MEF2 from a relatively uniform distribution through the nucleus into punctate nuclear bodies, where it colocalized with HDAC4. As MEF2 has also been implicated in regulation of long-term memory, these data suggest that the repressive effects of HDAC4 on long-term memory may be through interaction with MEF2. In the same genetic background, we also found that RNAi-mediated knockdown of HDAC4 impairs long-term memory, therefore we demonstrate that HDAC4 is not only a repressor of long-term memory, but also modulates normal memory formation.}, number={12}, journal={PLoS ONE}, publisher={Public Library of Science (PLoS)}, author={Fitzsimons, Helen L. and Schwartz, Silvia and Given, Fiona M. and Scott, Maxwell J.}, editor={Skoulakis, Efthimios M. C.Editor}, year={2013}, month={Dec}, pages={e83903} } @article{sze_dunham_carey_chang_li_edman_fjeldsted_scott_nuzhdin_tarone_2012, title={A de novo transcriptome assembly of Lucilia sericata (Diptera: Calliphoridae) with predicted alternative splices, single nucleotide polymorphisms and transcript expression estimates}, volume={21}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1111/j.1365-2583.2011.01127.x}, DOI={10.1111/j.1365-2583.2011.01127.x}, abstractNote={AbstractThe blow flyLucilia sericata(Diptera: Calliphoridae) (Meigen) is a nonmodel organism with no reference genome that is associated with numerous areas of research spanning the ecological, evolutionary, medical, veterinary and forensic sciences. To facilitate scientific discovery in this species, the transcriptome was assembled from more than six billion bases of Illumina and twenty‐one million bases of 454 sequence derived from embryonic, larval, pupal, adult and larval salivary gland libraries. The assembly was carried out in a manner that enabled identification of putative single nucleotide polymorphisms (SNPs) and alternative splices, and that provided expression estimates for various life history stages and for salivary tissue. The assembled transcriptome was also used to identify transcribed transposable elements inL. sericata. The results of this study will enable blow fly biologists, dipterists and comparative genomicists to more rapidly develop and test molecular and genetic hypotheses, especially those regarding blow fly development and salivary gland biology.}, number={2}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Sze, S‐H. and Dunham, J. P. and Carey, B. and Chang, P. L. and Li, F. and Edman, R. M. and Fjeldsted, C. and Scott, M. J. and Nuzhdin, S. V. and Tarone, A. M.}, year={2012}, month={Jan}, pages={205–221} } @article{stasiuk_scott_grant_2012, title={Developmental plasticity and the evolution of parasitism in an unusual nematode, Parastrongyloides trichosuri}, volume={3}, ISSN={2041-9139}, url={http://dx.doi.org/10.1186/2041-9139-3-1}, DOI={10.1186/2041-9139-3-1}, abstractNote={Abstract Background Parasitism is an important life history strategy in many metazoan taxa. This is particularly true of the Phylum Nematoda, in which parasitism has evolved independently at least nine times. The apparent ease with which parasitism has evolved amongst nematodes may, in part, be due to a feature of nematode development acting as a pre-adaptation for the transition from a free-living to a parasitic life history. One candidate pre-adaptive feature for evolution in terrestrial nematodes is the dauer larva, a developmentally arrested morph formed in response to environmental signals. Results We investigated the role of dauer development in the nematode, Parastrongyloides trichosuri, which has retained a complete free-living life cycle in addition to a life cycle as a mammalian gastrointestinal parasite. We show that the developmental switch between these life histories is sensitive to the same environmental cues as dauer arrest in free-living nematodes, including sensitivity to a chemical cue produced by the free-living stages. Furthermore, we show that genetic variation for the sensitivity of the cue(s) exists in natural populations of P. trichosuri, such that we derived inbred lines that were largely insensitive to the cue and other lines that were supersensitive to the cue. Conclusions For this parasitic clade, and perhaps more widely in the phylum, the evolution of parasitism co-opted the dauer switch of a free-living ancestor. This lends direct support to the hypothesis that the switch to developmental arrest in the dauer larva acted as a pre-adaptation for the evolution of parasitism, and suggests that the sensory transduction machinery downstream of the cue may have been similarly co-opted and modified. }, number={1}, journal={EvoDevo}, publisher={Springer Science and Business Media LLC}, author={Stasiuk, Susan J and Scott, Maxwell J and Grant, Warwick N}, year={2012}, month={Jan} } @article{concha_edman_belikoff_schiemann_carey_scott_2012, title={Organization and expression of the Australian sheep blowfly (Lucilia cuprina) hsp23, hsp24, hsp70 and hsp83 genes}, volume={21}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1111/j.1365-2583.2011.01123.x}, DOI={10.1111/j.1365-2583.2011.01123.x}, abstractNote={AbstractIn this study we report the isolation and characterization of a heat shock protein 70 (hsp70) gene, the hsp83 gene and two genes that encode small Hsps (Lchsp23 and Lchsp24) from the Australian sheep blowfly, Lucilia cuprina, a major agricultural pest. Phylogenetic analyses indicate that the LcHsp23 protein is the orthologue of Drosophila melanogaster Hsp23 and LcHsp24 is the orthologue of Sarcophaga crassipalpis Hsp23. Quantitative reverse‐transcriptase PCR analysis showed that the basal level of Lchsp83 RNA is relatively high at all developmental stages and only moderately induced by heat shock. In contrast, Lchsp70 transcripts are present at low levels and strongly induced by heat shock at all stages. The basal levels of expression and degrees of heat induction of the Lchsp23 and Lchsp24 transcripts were more variable across the different developmental stages. Putative heat shock factor binding sites were identified in the Lchsp24, Lchsp70 and Lchsp83 gene promoters. The isolation of these hsp gene promoters will facilitate constitutive or conditional expression of a gene of interest in transgenic Lucilia.}, number={2}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Concha, C. and Edman, R. M. and Belikoff, E. J. and Schiemann, A. H. and Carey, B. and Scott, M. J.}, year={2012}, month={Jan}, pages={169–180} } @article{laverty_li_belikoff_scott_2011, title={Abnormal Dosage Compensation of Reporter Genes Driven by the Drosophila Glass Multiple Reporter (GMR) Enhancer-Promoter}, volume={6}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0020455}, DOI={10.1371/journal.pone.0020455}, abstractNote={In Drosophila melanogaster the male specific lethal (MSL) complex is required for upregulation of expression of most X-linked genes in males, thereby achieving X chromosome dosage compensation. The MSL complex is highly enriched across most active X-linked genes with a bias towards the 3′ end. Previous studies have shown that gene transcription facilitates MSL complex binding but the type of promoter did not appear to be important. We have made the surprising observation that genes driven by the glass multiple reporter (GMR) enhancer-promoter are not dosage compensated at X-linked sites. The GMR promoter is active in all cells in, and posterior to, the morphogenetic furrow of the developing eye disc. Using phiC31 integrase-mediated targeted integration, we measured expression of lacZ reporter genes driven by either the GMR or armadillo (arm) promoters at each of three X-linked sites. At all sites, the arm-lacZ reporter gene was dosage compensated but GMR-lacZ was not. We have investigated why GMR-driven genes are not dosage compensated. Earlier or constitutive expression of GMR-lacZ did not affect the level of compensation. Neither did proximity to a strong MSL binding site. However, replacement of the hsp70 minimal promoter with a minimal promoter from the X-linked 6-Phosphogluconate dehydrogenase gene did restore partial dosage compensation. Similarly, insertion of binding sites for the GAGA and DREF factors upstream of the GMR promoter led to significantly higher lacZ expression in males than females. GAGA and DREF have been implicated to play a role in dosage compensation. We conclude that the gene promoter can affect MSL complex-mediated upregulation and dosage compensation. Further, it appears that the nature of the basal promoter and the presence of binding sites for specific factors influence the ability of a gene promoter to respond to the MSL complex.}, number={5}, journal={PLoS ONE}, publisher={Public Library of Science (PLoS)}, author={Laverty, Corey and Li, Fang and Belikoff, Esther J. and Scott, Maxwell J.}, editor={Bergmann, AndreasEditor}, year={2011}, month={May}, pages={e20455} } @article{fitzsimons_scott_2011, title={Genetic Modulation of Rpd3 Expression Impairs Long-Term Courtship Memory in Drosophila}, volume={6}, ISSN={1932-6203}, url={http://dx.doi.org/10.1371/journal.pone.0029171}, DOI={10.1371/journal.pone.0029171}, abstractNote={There is increasing evidence that regulation of local chromatin structure is a critical mechanism underlying the consolidation of long-term memory (LTM), however considerably less is understood about the specific mechanisms by which these epigenetic effects are mediated. Furthermore, the importance of histone acetylation in Drosophila memory has not been reported. The histone deacetylase (HDAC) Rpd3 is abundant in the adult fly brain, suggesting a post-mitotic function. Here, we investigated the role of Rpd3 in long-term courtship memory in Drosophila. We found that while modulation of Rpd3 levels predominantly in the adult mushroom body had no observed impact on immediate recall or one-hour memory, 24-hour LTM was severely impaired. Surprisingly, both overexpression as well as RNAi-mediated knockdown of Rpd3 resulted in impairment of long-term courtship memory, suggesting that the dose of Rpd3 is critical for normal LTM.}, number={12}, journal={PLoS ONE}, publisher={Public Library of Science (PLoS)}, author={Fitzsimons, Helen L. and Scott, Maxwell J.}, editor={Skoulakis, Efthimios M. C.Editor}, year={2011}, month={Dec}, pages={e29171} } @article{concha_li_scott_2010, title={Conservation and sex-specific splicing of the doublesex gene in the economically important pest species Lucilia cuprina}, volume={89}, ISSN={0022-1333 0973-7731}, url={http://dx.doi.org/10.1007/s12041-010-0039-5}, DOI={10.1007/s12041-010-0039-5}, abstractNote={Genetic control of sex determination in insects has been best characterized in Drosophila melanogaster, where the master gene Sxl codes for RNA that is sex specifically spliced to produce a functional protein only in females. SXL regulates the sex-specific splicing of transformer (tra) RNA which, in turn, regulates the splicing of dsx RNA to produce functional male and female proteins. In the Australian sheep blowfly (Lucilia cuprina), the tra gene (Lctra) is required for female development and Lctra transcripts are sex-specifically spliced such that only female Lctra mRNA codes for functional protein. In males, a factor encoded by the Y-linked male determining gene is thought to prevent the female-mode of splicing of Lctra RNA. To further our understanding of the sex determination regulatory hierarchy in L. cuprina, we have isolated the dsx gene (Lcdsx) from this species. We found that the Lcdsx transcripts are sex-specifically spliced in a similar manner as their counterparts in D. melanogaster, housefly and tephritids. The LcDSX proteins are well conserved and the male form of DSX contains a motif encoded by a male-specific exon that is within the female-specific intron. This intron/exon arrangement had previously been found only in the housefly dsx gene, suggesting this may be a unique feature of dsx genes of Calyptratae species.}, number={3}, journal={Journal of Genetics}, publisher={Springer Science and Business Media LLC}, author={Concha, Carolina and Li, Fang and Scott, Maxwell J.}, year={2010}, month={Sep}, pages={279–285} } @article{concha_belikoff_carey_li_schiemann_scott_2011, title={Efficient germ-line transformation of the economically important pest species Lucilia cuprina and Lucilia sericata (Diptera, Calliphoridae)}, volume={41}, ISSN={0965-1748}, url={http://dx.doi.org/10.1016/j.ibmb.2010.09.006}, DOI={10.1016/j.ibmb.2010.09.006}, abstractNote={The green blowfly species Lucilia cuprina and Lucilia sericata are economically important pests for the sheep industries of Australia and New Zealand. L. cuprina has long been considered a good target for a genetic pest management program. In addition, L. sericata maggots are used in the cleaning of wounds and necrotic tissue of patients suffering from ulcers that are difficult to treat by other methods. Development of efficient transgenesis methods would greatly facilitate the development of strains ideal for genetic control programs or could potentially improve "maggot therapy". We have previously reported the germ-line transformation of L. cuprina and the design of a "female killing system" that could potentially be applied to this species. However, the efficiency of transformation obtained was low and transformed lines were difficult to detect due to the low expression of the EGFP marker used. Here we describe an efficient and reliable method for germ-line transformation of L. cuprina using new piggyBac vector and helper plasmids containing the strong promoter from the L. cuprina hsp83 gene to drive expression of the transposase and fluorescent protein marker gene. We also report, for the first time, the germ-line transformation of L. sericata using the new piggyBac vector/helper combination.}, number={1}, journal={Insect Biochemistry and Molecular Biology}, publisher={Elsevier BV}, author={Concha, Carolina and Belikoff, Esther J. and Carey, Brandi-lee and Li, Fang and Schiemann, Anja H. and Scott, Maxwell J.}, year={2011}, month={Jan}, pages={70–75} } @article{scott_atapattu_schiemann_concha_henry_carey_belikoff_heinrich_sarkar_2010, title={Organisation and expression of a cluster of yolk protein genes in the Australian sheep blowfly, Lucilia cuprina}, volume={139}, ISSN={0016-6707 1573-6857}, url={http://dx.doi.org/10.1007/s10709-010-9492-6}, DOI={10.1007/s10709-010-9492-6}, abstractNote={The Australian sheep blowfly Lucilia cuprina is a major pest for the Australian and New Zealand sheep industries. With the long-term aim of making a strain of L. cuprina suitable for a genetic control program, we previously developed a tetracycline-repressible female lethal genetic system in Drosophila. A key part of this system is a female-specific promoter from a yolk protein (yp) gene controlling expression of the tetracycline-dependent transactivator (tTA). Here we report the sequence of a 14.2 kb genomic clone from L. cuprina that contains a cluster of three complete yp genes and one partial yp gene. The Lcyp genes are specifically expressed in females that have received a protein meal. A bioinformatic analysis of the promoter of one of the yp genes (LcypA) identified several putative binding sites for DSX, a known regulator of yp gene expression in other Diptera. A transgenic strain of L. cuprina was made that contained the LcypA promoter driving the expression of the Escherichia coli lacZ reporter gene. Transgenic females express high levels of β-galactosidase after a protein meal. Thus the LcypA promoter could be used to obtain female-specific expression of tTA in transgenic L. cuprina.}, number={1}, journal={Genetica}, publisher={Springer Science and Business Media LLC}, author={Scott, Maxwell J. and Atapattu, Asela and Schiemann, Anja H. and Concha, Carolina and Henry, Rebecca and Carey, Brandi-lee and Belikoff, Esther J. and Heinrich, Jörg C. and Sarkar, Abhimanyu}, year={2010}, month={Sep}, pages={63–70} } @article{schiemann_li_weake_belikoff_klemmer_moore_scott_2010, title={Sex-biased transcription enhancement by a 5' tethered Gal4-MOF histone acetyltransferase fusion protein in Drosophila}, volume={11}, ISSN={1471-2199}, url={http://dx.doi.org/10.1186/1471-2199-11-80}, DOI={10.1186/1471-2199-11-80}, abstractNote={Abstract Background In male Drosophila melanogaster, the male specific lethal (MSL) complex is somehow responsible for a two-fold increase in transcription of most X-linked genes, which are enriched for histone H4 acetylated at lysine 16 (H4K16ac). This acetylation requires MOF, a histone acetyltransferase that is a component of the MSL complex. MOF also associates with the non-specific lethal or NSL complex. The MSL complex is bound within active genes on the male X chromosome with a 3' bias. In contrast, the NSL complex is enriched at promoter regions of many autosomal and X-linked genes in both sexes. In this study we have investigated the role of MOF as a transcriptional activator. Results MOF was fused to the DNA binding domain of Gal4 and targeted to the promoter region of UAS-reporter genes in Drosophila. We found that expression of a UAS-red fluorescent protein (DsRed) reporter gene was strongly induced by Gal4-MOF. However, DsRed RNA levels were about seven times higher in female than male larvae. Immunostaining of polytene chromosomes showed that Gal4-MOF co-localized with MSL1 to many sites on the X chromosome in male but not female nuclei. However, in female nuclei that express MSL2, Gal4-MOF co-localized with MSL1 to many sites on polytene chromosomes but DsRed expression was reduced. Mutation of conserved active site residues in MOF (Glu714 and Cys680) reduced HAT activity in vitro and UAS-DsRed activation in Drosophila. In the presence of Gal4-MOF, H4K16ac levels were enriched over UAS-lacZ and UAS-arm-lacZ reporter genes. The latter utilizes the constitutive promoter from the arm gene to drive lacZ expression. In contrast to the strong induction of UAS-DsRed expression, UAS-arm-lacZ expression increased by about 2-fold in both sexes. Conclusions Targeting MOF to reporter genes led to transcription enhancement and acetylation of histone H4 at lysine 16. Histone acetyltransferase activity was required for the full transcriptional response. Incorporation of Gal4-MOF into the MSL complex in males led to a lower transcription enhancement of UAS-DsRed but not UAS-arm-lacZ genes. We discuss how association of Gal4-MOF with the MSL or NSL proteins could explain our results. }, number={1}, journal={BMC Molecular Biology}, publisher={Springer Science and Business Media LLC}, author={Schiemann, Anja H and Li, Fang and Weake, Vikki M and Belikoff, Esther J and Klemmer, Kent C and Moore, Stanley A and Scott, Maxwell J}, year={2010}, month={Nov} } @article{moore_ferhatoglu_jia_al-jiab_scott_2010, title={Structural and biochemical studies on the chromo-barrel domain of male specific lethal 3 (MSL3) reveal a binding preference for mono-or dimethyllysine 20 on histone H4}, volume={285}, number={52}, journal={Journal of Biological Chemistry}, publisher={American Society for Biochemistry and Molecular Biology}, author={Moore, Stanley A and Ferhatoglu, Yurdagul and Jia, Yunhua and Al-Jiab, Rami A and Scott, Maxwell J}, year={2010}, pages={40879–40890} } @article{schiemann_weake_li_laverty_belikoff_scott_2010, title={The importance of location and orientation of male specific lethal complex binding sites of differing affinities on reporter gene dosage compensation in Drosophila}, volume={402}, ISSN={0006-291X}, url={http://dx.doi.org/10.1016/j.bbrc.2010.10.088}, DOI={10.1016/j.bbrc.2010.10.088}, abstractNote={The male specific lethal (MSL) complex is required for X chromosome dosage compensation in Drosophila. The complex binds to most actively transcribed X-linked genes in males and upregulates expression. High resolution chromatin immunoprecipitation assays have identified over one hundred high affinity binding sites on the X chromosome. One of the first high affinity sites discovered is at cytological location 18D11. The MSL complex binds weakly to a single copy of a 510bp fragment from 18D11 but strongly to a tetramer of the fragment. Here we have investigated the effect of insertion of sites of differing affinities, either upstream or within the transcribed gene, on complex binding and transcription upregulation. Insertion of four copies of the 18D11 fragment upstream or at the 3' end of a reporter gene led to strong MSL complex binding and increased expression in males. In contrast, the MSL complex did not bind consistently to autosomal transgenes that contained a single copy of the 18D11 site upstream of the gene promoter. However, MSL complex binding was observed in all lines if the single 18D11 fragment was inserted into the 3' end of the reporter gene in either orientation. This is consistent with previous studies that showed gene transcription facilitates MSL complex binding. Surprisingly, transcription elevation in males was only observed if the 18D11 fragment was in the forward orientation and only in some lines. Our results suggest that MSL complex binding to weaker sites and transcription enhancement is influenced by gene transcription, binding site orientation and the local chromatin environment. In contrast, strong binding sites do not need to be transcribed to recruit sufficient complex to cause transcription elevation of nearby genes.}, number={4}, journal={Biochemical and Biophysical Research Communications}, publisher={Elsevier BV}, author={Schiemann, Anja H. and Weake, Vikki M. and Li, Fang and Laverty, Corey and Belikoff, Esther J. and Scott, Maxwell J.}, year={2010}, month={Nov}, pages={699–704} } @article{concha_scott_2009, title={Sexual Development in Lucilia cuprina (Diptera, Calliphoridae) Is Controlled by the Transformer Gene}, volume={182}, ISSN={1943-2631}, url={http://dx.doi.org/10.1534/genetics.109.100982}, DOI={10.1534/genetics.109.100982}, abstractNote={Abstract Insects use an amazing variety of genetic systems to control sexual development. A Y-linked male determining gene (M) controls sex in the Australian sheep blowfly Lucilia cuprina, an important pest insect. In this study, we isolated the L. cuprina transformer (Lctra) and transformer2 (Lctra2) genes, which are potential targets of M. The LCTRA and LCTRA2 proteins are significantly more similar to homologs from tephritid insects than Drosophila. The Lctra transcript is alternatively spliced such that only females make a full-length protein and the presence of six TRA/TRA2 binding sites in the female first intron suggest that Lctra splicing is autoregulated as in tephritids. LCTRA is essential for female development as RNAi knockdown of Lctra mRNA leads to the development of male genitalia in XX adults. Analysis of Lctra expression during development shows that early and midstage male and female embryos express the female form of Lctra and males express only the male form by the first instar larval stage. Our results suggest that an autoregulatory loop sustains female development and that expression of M inhibits Lctra autoregulation, switching its splicing to the male form. The conservation of tra function and regulation in a Calliphorid insect shows that this sex determination system is not confined to Tephritidae. Isolation of these genes is an important step toward the development of a strain of L. cuprina suitable for a genetic control program.}, number={3}, journal={Genetics}, publisher={Oxford University Press (OUP)}, author={Concha, Carolina and Scott, Maxwell J}, year={2009}, month={Jul}, pages={785–798} } @article{concha_scott_2009, title={Sexual development in Lucilia cuprina (Diptera, Calliphoridae) is controlled by the transformer gene}, volume={182}, number={3}, journal={Genetics}, publisher={Genetics Society of America}, author={Concha, Carolina and Scott, Maxwell J}, year={2009}, pages={785–798} } @article{scott_li_2008, title={How do ncRNAs guide chromatin-modifying complexes to specific locations within the nucleus?}, volume={5}, ISSN={1547-6286 1555-8584}, url={http://dx.doi.org/10.4161/rna.5.1.5943}, DOI={10.4161/rna.5.1.5943}, abstractNote={Transcriptome analyses have led to the realisation that eukaryotic cells make a large number of noncoding RNAs (ncRNAs). It appears that some of these are involved in guiding chromatin-modifying complexes to specific locations within the nucleus. How such ncRNAs function is largely unknown but various models have been proposed. Here we briefly discuss the evidence supporting two such models; that ncRNAs function by annealing either with nascent transcripts or with homologous DNA sequences. We then review a third model that is based on our recent work on the role of the noncoding roX RNAs in the localisation of the MSL complex to sites on the X chromosome in Drosophila. Our results suggest that the MSL1 and MSL2 proteins bind to chromatin but it is the incorporation of the roX RNAs into the complex that somehow alters the binding specificity of the MSL1/MSL2 proteins to recognise sites on the X chromosome.}, number={1}, journal={RNA Biology}, publisher={Informa UK Limited}, author={Scott, Maxwell J. and Li, Fang}, year={2008}, month={Jan}, pages={13–16} } @article{li_schiemann_scott_2008, title={Incorporation of the Noncoding roX RNAs Alters the Chromatin-Binding Specificity of the Drosophila MSL1/MSL2 Complex}, volume={28}, ISSN={1098-5549}, url={http://dx.doi.org/10.1128/MCB.00309-08}, DOI={10.1128/MCB.00309-08}, abstractNote={This article refers to:Incorporation of the Noncoding roX RNAs Alters the Chromatin-Binding Specificity of the Drosophila MSL1/MSL2 Complex}, number={8}, journal={Molecular and Cellular Biology}, publisher={Informa UK Limited}, author={Li, F> and Schiemann, A.H. and Scott, M.J}, year={2008}, month={Apr}, pages={2850–2850} } @article{weake_scott_2007, title={The non-dosage compensated Lsp1α gene of Drosophila melanogaster escapes acetylation by MOF in larval fat body nuclei, but is flanked by two dosage compensated genes}, volume={8}, ISSN={1471-2199}, url={http://dx.doi.org/10.1186/1471-2199-8-35}, DOI={10.1186/1471-2199-8-35}, abstractNote={In Drosophila melanogaster dosage compensation of most X-linked genes is mediated by the male-specific lethal (MSL) complex, which includes MOF. MOF acetylates histone H4 at lysine 16 (H4K16ac). The X-linked Larval serum protein one α (Lsp1 α) gene has long been known to be not dosage compensated. Here we have examined possible explanations for why the Lsp1 α gene is not dosage compensated. Quantitative RNase protection analysis showed that the genes flanking Lsp1 α are expressed equally in males and females and confirmed that Lsp1 α is not dosage compensated. Unlike control X-linked genes, Lsp1 α was not enriched for H4K16ac in the third instar larval fat body, the tissue in which the gene is actively expressed. X-linked Lsp1α promoter-lacZ reporter transgenes are enriched for H4K16ac in third instar larval fat body. An X-linked reporter gene bracketed by Lsp1 α flanking regions was dosage compensated. One of the genes flanking Lsp1 α is expressed in the same tissue. This gene shows a modest enrichment for H4K16ac but only at the part of the gene most distant from Lsp1 α. Phylogenetic analyses of the sequences of the genomes of 12 Drosophila species shows that Lsp1 α is only present within the melanogaster subgroup of species. Lsp1 α is not modified by the MSL complex but is in a region of the X chromosome that is regulated by the MSL complex. The high activity or tissue-specificity of the Lsp1 α promoter does not prevent regulation by the MSL complex. The regions flanking Lsp1 α do not appear to block access by the MSL complex. Lsp1 α appears to have recently evolved within the melanogaster subgroup of Drosophila species. The most likely explanation for why Lsp1 α is not dosage compensated is that the gene has not evolved a mechanism to independently recruit the MSL complex, possibly because of its recent evolutionary origin, and because there appears to be a low level of bound MSL complex in a nearby gene that is active in the same tissue.}, number={1}, journal={BMC Molecular Biology}, publisher={Springer Science and Business Media LLC}, author={Weake, Vikki M and Scott, Maxwell J}, year={2007}, pages={35} } @article{sarkar_atapattu_belikoff_heinrich_li_horn_wimmer_scott_2006, title={Insulated piggyBac vectors for insect transgenesis}, volume={6}, ISSN={1472-6750}, url={http://dx.doi.org/10.1186/1472-6750-6-27}, DOI={10.1186/1472-6750-6-27}, abstractNote={Abstract Background Germ-line transformation of insects is now a widely used method for analyzing gene function and for the development of genetically modified strains suitable for pest control programs. The most widely used transposable element for the germ-line transformation of insects is piggyBac. The site of integration of the transgene can influence gene expression due to the effects of nearby transcription enhancers or silent heterochromatic regions. Position effects can be minimized by flanking a transgene with insulator elements. The scs/scs' and gypsy insulators from Drosophila melanogaster as well as the chicken β-globin HS4 insulator function in both Drosophila and mammalian cells. Results To minimize position effects we have created a set of piggyBac transformation vectors that contain either the scs/scs', gypsy or chicken β-globin HS4 insulators. The vectors contain either fluorescent protein or eye color marker genes and have been successfully used for germ-line transformation of Drosophila melanogaster. A set of the scs/scs' vectors contains the coral reef fluorescent protein marker genes AmCyan, ZsGreen and DsRed that have not been optimized for translation in human cells. These marker genes are controlled by a combined GMR-3xP3 enhancer/promoter that gives particularly strong expression in the eyes. This is also the first report of the use of the ZsGreen and AmCyan reef fluorescent proteins as transformation markers in insects. Conclusion The insulated piggyBac vectors should protect transgenes against position effects and thus facilitate fine control of gene expression in a wide spectrum of insect species. These vectors may also be used for transgenesis in other invertebrate species. }, number={1}, journal={BMC Biotechnology}, publisher={Springer Science and Business Media LLC}, author={Sarkar, Abhimanyu and Atapattu, Asela and Belikoff, Esther J and Heinrich, Jörg C and Li, Xuelei and Horn, Carsten and Wimmer, Ernst A and Scott, Maxwell J}, year={2006}, month={Jun}, pages={27} } @article{scott_sarkar_belikoff_2005, title={Organisation and Expression of a Cluster of Female-specific Genes in the Australian Sheep Blowfly, Lucilia cuprina}, journal={Area-Wide Control of Insect Pests}, author={Scott, MJ and Sarkar, A and Belikoff, EJ}, year={2005} } @article{li_parry_scott_2005, title={The Amino-Terminal Region of Drosophila MSL1 Contains Basic, Glycine-Rich, and Leucine Zipper-Like Motifs That Promote X Chromosome Binding, Self-Association, and MSL2 Binding, Respectively}, volume={25}, ISSN={1098-5549}, url={http://dx.doi.org/10.1128/MCB.25.20.8913-8924.2005}, DOI={10.1128/MCB.25.20.8913-8924.2005}, abstractNote={ABSTRACT In Drosophila melanogaster, X chromosome dosage compensation is achieved by doubling the transcription of most X-linked genes. The male-specific lethal (MSL) complex is required for this process and binds to hundreds of sites on the male X chromosome. The MSL1 protein is essential for X chromosome binding and serves as a central scaffold for MSL complex assembly. We find that the amino-terminal region of MSL1 binds to hundreds of sites on the X chromosome in normal males but only to approximately 30 high-affinity sites in the absence of endogenous MSL1. Binding to the high-affinity sites requires a basic motif at the amino terminus that is conserved among Drosophila species. X chromosome binding also requires a conserved leucine zipper-like motif that binds to MSL2. A glycine-rich motif between the basic and leucine-zipper-like motifs mediates MSL1 self-association in vitro and binding of the amino-terminal region of MSL1 to the MSL complex assembled on the male X chromosome. We propose that the basic region may mediate DNA binding and that the glycine-rich region may promote the association of MSL complexes to closely adjacent sites on the X chromosome.}, number={20}, journal={Molecular and Cellular Biology}, publisher={Informa UK Limited}, author={Li, Fang and Parry, David A. D. and Scott, Maxwell J.}, year={2005}, month={Oct}, pages={8913–8924} } @article{scott_heinrich_li_2004, title={Progress towards the development of a transgenic strain of the Australian sheep blowfly (Lucilia cuprina) suitable for a male-only sterile release program}, volume={34}, number={2}, journal={Insect biochemistry and molecular biology}, publisher={Pergamon}, author={Scott, Maxwell J and Heinrich, Jörg C and Li, Xuelei}, year={2004}, pages={185–192} } @article{heinrich_li_henry_haack_stringfellow_heath_scott_2002, title={Germ‐line transformation of the Australian sheep blowfly Lucilia cuprina}, volume={11}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1046/j.0962-1075.2001.00301.x}, DOI={10.1046/j.0962-1075.2001.00301.x}, abstractNote={AbstractThe Australian sheep blowfly, Lucilia cuprina, is the most important economic insect pest for the sheep industries in Australia and New Zealand. piggyBac‐mediated germ‐line transformation of L. cuprina was achieved with a helper plasmid that had the Drosophila melanogaster hsp70 promoter controlling expression of the transposase and a piggyBac vector with an EGFP marker gene. Two transformant lines were obtained, at a frequency of approximately 1–2% per fertile G0. One of these lines has a single copy of the transgene, the other most likely has four copies. This is the first report of germ‐line transformation of L. cuprina and is an important step towards the generation of engineered strains that would be suitable for male‐only release eradication/suppression programmes.}, number={1}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Heinrich, J. C. and Li, X. and Henry, R. A. and Haack, N. and Stringfellow, L. and Heath, A. C. G. and Scott, M. J.}, year={2002}, month={Feb}, pages={1–10} } @article{scott_2001, title={Making 1X=2X: Studies on dosage compensation in Drosophila}, volume={9}, number={4}, journal={NZ BioScience}, author={Scott, M.J.}, year={2001}, pages={23–24} } @article{scott_2001, title={Progress towards the development of a transgenic strain of the Australian sheep blowfly suitable for a sterile-release program}, volume={9}, number={2}, journal={NZ BioScience}, author={Scott, M.J}, year={2001}, pages={11–13} } @article{henry_tews_li_scott_2001, title={Recruitment of the Male-specific Lethal (MSL) Dosage Compensation Complex to an Autosomally Integrated roXChromatin Entry Site Correlates with an Increased Expression of an Adjacent Reporter Gene in Male Drosophila}, volume={276}, ISSN={0021-9258}, url={http://dx.doi.org/10.1074/jbc.M103008200}, DOI={10.1074/jbc.M103008200}, abstractNote={Drosophila dosage compensate (equalize X-linked gene products) by doubling the transcription of most X-linked genes in males. The MSL (male-specific lethal) ribonucleoprotein complex consisting of at least five proteins and two non-coding RNAs (roX1 and roX2) is essential for this transcription response. Recently it has been shown that the X-linked roX1 and roX2 genes each contain at least one chromatin entry site for the MSL complex. In this study we show that insertion of either roX1 or roX2 DNA sequences, upstream of an insulated lacZ reporter gene controlled with the constitutive armadillo promoter (arm-lacZ), results in a significant elevation of expression of lacZ in males. However, full compensation, that is a precise doubling of lacZ expression in males relative to females, was only observed in some lines carrying autosomal insertions of either roX1-arm-lacZ orroX2-arm-lacZ transgenes. Furthermore, we found that a 419-base pair fragment of roX1 that contains an MSL binding site was sufficient to cause a modest elevation of expression oflacZ in males, but this response was significantly less than obtained with a full-length roX1 cDNA. This is the first direct demonstration that insertion of an MSL chromatin entry site on an autosome results in elevated expression in males of genes near the entry site.}, number={34}, journal={Journal of Biological Chemistry}, publisher={Elsevier BV}, author={Henry, Rebecca A. and Tews, Birke and Li, Xuelei and Scott, Maxwell J.}, year={2001}, month={Aug}, pages={31953–31958} } @article{li_heinrich_scott_2001, title={piggyBac-mediated transposition in Drosophila melanogaster: an evaluation of the use of constitutive promoters to control transposase gene expression}, volume={10}, number={5}, journal={Insect molecular biology}, publisher={Blackwell Science Ltd}, author={Li, X and Heinrich, JC and Scott, MJ}, year={2001}, pages={447–455} } @article{li_heinrich_scott_2001, title={piggyBac‐mediated transposition in Drosophila melanogaster: an evaluation of the use of constitutive promoters to control transposase gene expression}, volume={10}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1046/j.0962-1075.2001.00283.x}, DOI={10.1046/j.0962-1075.2001.00283.x}, abstractNote={AbstractTransgenic non‐Drosophilid insects have been made using insect transposable elements that have a broad host range such as the piggyBac element. However, the success rate is often low. Previous piggyBac helper plasmids have used either the piggyBac or the hsp70 promoter from Drosophila melanogaster to control expression of the transposase gene. Here we show that plasmids with the piggyBac transposase gene regulated by constitutive promoters can be effective ‘helpers’ for mediating transposition in D. melanogaster. We also present preliminary evidence on the use of an RNA helper that encodes the transposase. Our results suggest that for germ‐line transformation of non‐Drosophilid insects it may be advantageous to isolate a constitutive promoter from the species of interest to control transposase expression.}, number={5}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Li, X. and Heinrich, J. C. and Scott, M. J.}, year={2001}, month={Oct}, pages={447–455} } @article{heinrich_scott_2000, title={A repressible female-specific lethal genetic system for making transgenic insect strains suitable for a sterile-release program}, volume={97}, ISSN={0027-8424 1091-6490}, url={http://dx.doi.org/10.1073/pnas.140142697}, DOI={10.1073/pnas.140142697}, abstractNote={ We have developed a tetracycline-repressible female-specific lethal genetic system in the vinegar fly Drosophila melanogaster . One component of the system is the tetracycline-controlled transactivator gene under the control of the fat body and female-specific transcription enhancer from the yolk protein 1 gene. The other component consists of the proapoptotic gene hid under the control of a tetracycline-responsive element. Males and females of a strain carrying both components are viable on medium supplemented with tetracycline, but only males survive on normal medium. A strain with such properties would be ideal for a sterile-insect release program, which is most effective when only males are released in the field. }, number={15}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Heinrich, Jörg C. and Scott, Maxwell J.}, year={2000}, month={Jul}, pages={8229–8232} } @article{scott_2000, title={MSL1 plays a central role in assembly of the MSL complex, essential for dosage compensation in Drosophila}, volume={19}, ISSN={1460-2075}, url={http://dx.doi.org/10.1093/emboj/19.1.144}, DOI={10.1093/emboj/19.1.144}, abstractNote={In male Drosophila, histone H4 acetylated at Lys16 is enriched on the X chromosome, and most X‐linked genes are transcribed at a higher rate than in females (thus achieving dosage compensation). Five proteins, collectively called the MSLs, are required for dosage compensation and male viability. Here we show that one of these proteins, MSL1, interacts with three others, MSL2, MSL3 and MOF. The latter is a putative histone acetyl transferase. Overexpression of either the N‐ or C‐terminal domain of MSL1 has dominant‐negative effects, i.e. causes male‐specific lethality. The lethality due to expression of the N‐terminal domain is reduced if msl2 is co‐overexpressed. MSL2 co‐purifies over a FLAG affinity column with the tagged region of MSL1, and both MSL3 and MOF co‐purify with the FLAG‐tagged MSL1 C‐terminal domain. Furthermore, the MSL1 C‐terminal domain binds specifically to a GST–MOF fusion protein and co‐immunoprecipitates with HA‐tagged MSL3. The MSL1 C‐terminal domain shows similarity to a region of mouse CBP, a transcription co‐activator. We conclude that a main role of MSL1 is to serve as the backbone for assembly of the MSL complex.}, number={1}, journal={The EMBO Journal}, publisher={Springer Science and Business Media LLC}, author={Scott, M. J.}, year={2000}, month={Jan}, pages={144–155} } @article{scott_pan_cleland_knox_heinrich_2000, title={MSL1 plays a central role in assembly of the MSL complex, essential for dosage compensation in Drosophila}, volume={19}, number={1}, journal={The EMBO journal}, publisher={EMBO Press}, author={Scott, Maxwell J and Pan, Lewis L and Cleland, Sheralee B and Knox, Andrea L and Heinrich, Jörg}, year={2000}, pages={144–155} } @article{fitzsimons_henry_scott_1999, volume={105}, ISSN={0016-6707}, url={http://dx.doi.org/10.1023/a:1003801402153}, DOI={10.1023/a:1003801402153}, number={3}, journal={Genetica}, publisher={Springer Science and Business Media LLC}, author={Fitzsimons, Helen L. and Henry, Rebecca A. and Scott, Maxwell J.}, year={1999}, pages={215–226} } @article{fitzsimons_henry_scott_1999, title={Development of an insulated reporter system to search for cis-acting DNA sequences required for dosage compensation in Drosophila}, volume={105}, number={3}, journal={Genetica}, publisher={Kluwer Academic Publishers}, author={Fitzsimons, Helen L and Henry, Rebecca A and Scott, Maxwell J}, year={1999}, pages={215–226} } @article{frith_scott_1999, title={Isolation and characterisation of the Drosophila Dror2 gene}, volume={7}, journal={NZ BioScience}, author={Frith, K.J. and Scott, M.J.}, year={1999}, pages={17–22} } @article{banks_robinson_kwiatowski_ayala_scott_kriticou_1995, title={A second superoxide dismutase gene in the medfly, Ceratitis capitata.}, volume={140}, ISSN={1943-2631}, url={http://dx.doi.org/10.1093/genetics/140.2.697}, DOI={10.1093/genetics/140.2.697}, abstractNote={Abstract We report the first case of two Cu/Zn Sod genes (ccSod1 and ccSod2) that have been cloned and sequenced from an insect, the medfly, Ceratitis capitata. Biochemical evidence suggested the presence of two Sod genes in the medfly. The two genes are isolated using different molecular strategies: ccSod1 via cross-hybridization to a genomic library using a heterologous probe and ccSod2 from cDNA using a homologous probe generated by PCR. Sequence analysis shows that ccSod1 and ccSod2 are different genes. The inferred amino sequences show that all essential residues of the active site are strictly conserved, which suggests both genes encode functional Cu/Zn superoxide dismutase (SOD). Phylogenetic analysis by the maximum parsimony method with bootstrap resampling of previously known Cu/Zn SOD reveals two monophyletic groups, vertebrates and insects. The position of ccSOD2 in this phylogeny is undefined with respect to dipteran ccSOD1, vertebrate, plant, fungal, and extracellular Cu/Zn SOD, which suggests that the duplication detected in Ceratitis is ancient, perhaps as old as the origins of the arthropod phylum in the Cambrian more than 500 million years ago. In situ hybridization to polytene chromosomes places the genes on different chromosomes, which is consistent with an ancient gene duplication.}, number={2}, journal={Genetics}, publisher={Oxford University Press (OUP)}, author={Banks, G K and Robinson, A S and Kwiatowski, J and Ayala, F J and Scott, M J and Kriticou, D}, year={1995}, month={Jun}, pages={697–702} } @article{zhou_yang_scott_pannuti_fehr_eisen_koonin_fouts_wrightsman_manning_et al._1995, title={Male-specific lethal 2, a dosage compensation gene of Drosophila, undergoes sex-specific regulation and encodes a protein with a RING finger and a metallothionein-like cysteine cluster.}, volume={14}, ISSN={0261-4189}, url={http://dx.doi.org/10.1002/j.1460-2075.1995.tb07288.x}, DOI={10.1002/j.1460-2075.1995.tb07288.x}, abstractNote={In Drosophila the equalization of X‐linked gene products between males and females, i.e. dosage compensation, is the result of a 2‐fold hypertranscription of most of these genes in males. At least four regulatory genes are required for this process. Three of these genes, maleless (mle), male‐specific lethal 1 (msl‐1) and male‐specific lethal 3 (msl‐3), have been cloned and their products have been shown to interact and to bind to numerous sites on the X chromosome of males, but not of females. Although binding to the X chromosome is negatively correlated with the function of the master regulatory gene Sex lethal (Sxl), the mechanisms that restrict this binding to males and to the X chromosome are not yet understood. We have cloned the last of the known autosomal genes involved in dosage compensation, male‐specific lethal 2 (msl‐2), and characterized its product. The encoded protein (MSL‐2) consists of 769 amino acid residues and has a RING finger (C3HC4 zinc finger) and a metallothionein‐like domain with eight conserved and two non‐conserved cysteines. In addition, it contains a positively and a negatively charged amino acid residue cluster and a coiled coil domain that may be involved in protein‐protein interactions. Males produce a msl‐2 transcript that is shorter than in females, due to differential splicing of an intron of 132 bases in the untranslated leader. Using an antiserum against MSL‐2 we have shown that the protein is expressed at a detectable level only in males, where it is physically associated with the X chromosome. Our observations suggest that MSL‐2 may be the target of the master regulatory gene Sxl and provide the basic elements of a working hypothesis on the function of MSL‐2 in mediating the 2‐fold increase in transcription that is characteristic of dosage compensation.}, number={12}, journal={The EMBO Journal}, publisher={Springer Science and Business Media LLC}, author={Zhou, Shubo and Yang, Youfeng and Scott, Maxwell J and Pannuti, Antonio and Fehr, Krista C and Eisen, Arri and Koonin, Eugene V and Fouts, David L and Wrightsman, Ruth and Manning, Jerry E and et al.}, year={1995}, month={Jun}, pages={2884–2895} } @article{scott_kriticou_robinson_1993, title={Isolation of cDNAs encoding 6‐phosphogluconate dehydrogenase and glucose‐6‐phosphate dehydrogenase from the mediterranean fruit fly Ceratitis capitata: correlating genetic and physical maps of chromosome 5}, volume={1}, ISSN={0962-1075 1365-2583}, url={http://dx.doi.org/10.1111/j.1365-2583.1993.tb00094.x}, DOI={10.1111/j.1365-2583.1993.tb00094.x}, abstractNote={AbstractWe have isolated and determined the nucleotide sequences for cDNA clones encoding glucose‐6‐phosphate dehydrogenase (G6PD) and 6‐phosphogluconate dehydrogenase (6PGD) from the medfly Ceratitis capitata. The derived amino acid sequences for G6PD and 6PGD are presented and compared with G6PDs and 6PGDs from other species. The codon usage of the cDNA clones has little bias with the notable exceptions of arginine, glycine and leucine.The chromosomal location of the genes for 6PGD and G6PD were determined by in situ hybridization to salivary gland polytene chromosomes. This localization orients a genetic map of enzymatic loci and illustrates a remarkable similarity in the intra chromosomal order of homologous genes between Drosophila melanogaster and medfly.}, number={4}, journal={Insect Molecular Biology}, publisher={Wiley}, author={Scott, M. J. and Kriticou, D. and Robinson, A. S.}, year={1993}, month={May}, pages={213–222} } @article{scott_lucchesi_1991, title={Structure and expression of the Drosophila melanogaster gene encoding 6-phosphogluconate dehydrogenase}, volume={109}, ISSN={0378-1119}, url={http://dx.doi.org/10.1016/0378-1119(91)90607-d}, DOI={10.1016/0378-1119(91)90607-d}, abstractNote={We have determined the nucleotide sequence and structure of Pgd+, the Drosophila melanogaster gene that encodes the enzyme, 6-phosphogluconate dehydrogenase (6PGD). The derived 481-amino acid sequence for D. melanogaster 6PGD is presented and compared with 6PGD sequences from other species. To characterize the cis-acting sequences necessary for expression of Pgd+, fragments containing this gene as well as Pgd+ promoter-lacZ fusions were introduced into the D. melanogaster germ line by P-element-mediated transformation. Our results indicate that the large second intron is critical for Pgd+ expression in adults. Only 421 bp of Pgd+ 5′-flanking DNA are necessary to direct expression in imaginal discs, gonads and gut of third-instar larvae. Sequences downstream from the transcription start point are necessary for expression in the larval fat body.}, number={2}, journal={Gene}, publisher={Elsevier BV}, author={Scott, Maxwell J. and Lucchesi, John C.}, year={1991}, month={Dec}, pages={177–183} } @inbook{stein_scott_o’malley_1990, title={Intervening sequences in molecular evolution}, booktitle={Intervening Sequences in Evolution and Development}, publisher={Oxford University Press}, author={Stein, Joseph P. and Scott, Maxwell J. and O’Malley, Bert W.}, year={1990}, pages={92–111} } @inproceedings{scott_tsai_omalley_1987, title={DNASE I SENSITIVITY OF THE OVOMUCOID-OVOINHIBITOR GENE-COMPLEX IN OVIDUCT NUCLEI AND RELATIVE LOCATION OF CR-1 REPETITIVE SEQUENCES}, booktitle={JOURNAL OF CELLULAR BIOCHEMISTRY}, author={SCOTT, MJ and TSAI, MJ and OMALLEY, BW}, year={1987}, pages={91–91} } @article{scott_tsai_o'malley_1987, title={Deoxyribonuclease I sensitivity of the ovomucoid-ovoinhibitor gene complex in oviduct nuclei and relative location of CR1 repetitive sequences}, volume={26}, ISSN={0006-2960 1520-4995}, url={http://dx.doi.org/10.1021/bi00395a037}, DOI={10.1021/bi00395a037}, abstractNote={The location of CR1 middle repetitive sequences within or near the boundaries of the ovalbumin DNase I sensitive domain has suggested that CR1 sequences may play a role in defining transition regions of DNase I sensitivity in hen oviduct nuclei. We have examined this apparent relationship of CR1 sequences and transitions of chromatin structure by determining the DNase I sensitivity in oviduct nuclei of a 47-kilobase region that contains five CR1 sequences and the transcribed ovomucoid and ovoinhibitor genes. We find that three of the CR1 sequences occur within a broad transition region of decreasing DNase I sensitivity downstream of the ovomucoid gene. Another CR1 is in a region of decreased DNase I sensitivity within the ovoinhibitor gene. The fifth CR1 sequence is in a DNase I sensitive region between the two genes but which is less sensitive to DNase I digestion than the region immediately upstream from the ovomucoid gene. Thus, the CR1 sequences occur within regions of reduced relative DNase I sensitivity, suggesting that CR1s could facilitate the formation of a chromatin conformation that is less sensitive to DNase I digestion. Unexpectedly, the noncoding strand of sequences within and immediately adjacent to the 5' end of the actively transcribed ovomucoid and ovalbumin genes was less sensitive to DNase I digestion than their respective coding strands.}, number={21}, journal={Biochemistry}, publisher={American Chemical Society (ACS)}, author={Scott, Maxwell J. and Tsai, Ming Jer and O'Malley, Bert W.}, year={1987}, month={Oct}, pages={6831–6840} } @article{scott_huckaby_kato_kohr_laskowski_tsai_o'malley_1987, title={Ovoinhibitor introns specify functional domains as in the related and linked ovomucoid gene.}, volume={262}, ISSN={0021-9258}, url={http://dx.doi.org/10.1016/S0021-9258(18)45659-1}, DOI={10.1016/S0021-9258(18)45659-1}, abstractNote={We have isolated cDNA clones and determined the gene structure of chicken ovoinhibitor, a seven domain Kazal serine proteinase inhibitor. Using RNA blot hybridization analysis, the gene was identified initially as a region 9-23 kilobases upstream of the gene for the related inhibitor ovomucoid. Ovoinhibitor RNA appears in oviduct and liver. cDNA clones were identified by screening an oviduct cDNA library with a nick-translated DNA restriction fragment which contained an exon of the gene. The mature protein sequence derived from a cDNa clone is in excellent agreement with that which we obtained from direct sequencing of purified ovoinhibitor. The protein-sequencing strategy is reported. The P1 amino acids of the Kazal domains are consistent with the known broad inhibitory specificity of ovoinhibitor. The gene is about 10.3 kilobases in length and consists of 16 exons. Each Kazal domain is encoded by two exons. Like ovomucoid, introns fall between the coding sequences of the ovoinhibitor domains, an arrangement which may have facilitated domain duplication. The intradomain intron occurs in an identical position in all of the ovoinhibitor and ovomucoid Kazal domains, suggesting that this intron was present in the primordial inhibitor gene. We discuss the location of the intradomain intron in relation to the known structure of four Kazal inhibitors and suggest a scheme for the evolution of the ovoinhibitor gene.}, number={12}, journal={Journal of Biological Chemistry}, publisher={Elsevier BV}, author={Scott, M.J. and Huckaby, C.S. and Kato, I. and Kohr, W.J. and Laskowski, M., Jr. and Tsai, M.J. and O'Malley, B.W.}, year={1987}, month={Apr}, pages={5899–5907} } @book{scott_1986, title={Higher order chromatin structure of the ovomucoid-ovoinhibitor gene complex}, publisher={Baylor College of Medicine}, author={Scott, Maxwell John}, year={1986} } @inbook{tsai_tsai_baez_simmen_scott_o'malley_1985, title={Expression of eukaryotic genes: Transcription and analysis}, booktitle={Laboratory Methods Manual for Hormone Action and Molecular Endocrinology}, publisher={Houston Biological Assoc. Inc}, author={Tsai, M.-J. and Tsai, S.Y. and Baez, M. and Simmen, F.A. and Scott, M.J. and O'Malley, B.W.}, editor={Schrader, W.T. and O'Malley, B.W.Editors}, year={1985}, pages={13 1–13 4} } @inbook{tsai_tsai_baez_simmen_scott_o'malley_1985, title={Expression of eukaryotic genes: Transcription and analysis}, booktitle={Laboratory Methods Manual for Hormone Action and Molecular Endocrinology}, publisher={Houston Biological Assoc. Inc}, author={Tsai, M.-J. and Tsai, S.Y. and Baez, M. and Simmen, F.A. and Scott, M.J. and O'Malley, B.W.}, editor={Schrader, W.T. and O'Malley, B.W.Editors}, year={1985}, pages={13 1–13 4} } @inbook{huckaby_ciejek_scott_alevy_tsai_o'malley_1985, place={New York}, title={Nuclear matrix: Relationship to DNase I sensitivity and transcriptional activity}, volume={26}, booktitle={UCLA Symposia "Nuclear Envelope Structure and RNA Maturation}, publisher={Alan R. Liss, Inc}, author={Huckaby, C.S. and Ciejek, E.M. and Scott, M.J. and Alevy, M.C. and Tsai, M.-J. and O'Malley, B.W.}, editor={Smuckler, E.A. and Clawson, G.A.Editors}, year={1985}, pages={87–97} } @article{the structural organization of the chicken calmodulin gene._1985, volume={260}, number={2}, journal={Journal of Biological Chemistry}, publisher={American Society for Biochemistry and Molecular Biology}, year={1985}, pages={907–912} }