@article{li_yamamoto_belikoff_berger_griffith_scott_2021, title={A conditional female lethal system for genetic suppression of the global fruit crop pest <scp><i>Drosophila suzukii}, volume={77}, ISSN={1526-498X 1526-4998}, url={http://dx.doi.org/10.1002/ps.6530}, DOI={10.1002/ps.6530}, abstractNote={BACKGROUND
Drosophila 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 control of D. suzukii.


RESULTS
In this study we have 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.


CONCLUSION
Our data suggests that the transgenic strain could provide an effective approach for control of this invasive pest of soft-skinned fruits. This article is protected by copyright. All rights reserved.}, 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{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{paulo_williamson_arp_li_sagel_skoda_sanchez-gallego_vasquez_quintero_pérez de león_et al._2019, title={Specific Gene Disruption in the Major Livestock Pests<i>Cochliomyia hominivorax</i>and<i>Lucilia cuprina</i>Using CRISPR/Cas9}, volume={9}, ISSN={2160-1836}, url={http://dx.doi.org/10.1534/g3.119.400544}, DOI={10.1534/g3.119.400544}, abstractNote={Abstract}, 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 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}, 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{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{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{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={Abstract}, 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{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{concha_edman_belikoff_schiemann_carey_scott_2012, title={Organization and expression of the Australian sheep blowfly (<i>Lucilia cuprina</i>) <i>hsp23</i>, <i>hsp24, hsp70</i> and <i>hsp83</i> 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={Abstract}, 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{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}, 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} }