@article{yadav_butler_yamamoto_patil_lloyd_scott_2023, title={CRISPR/Cas9-based split homing gene drive targeting
            <i>doublesex
            for population suppression of the global fruit pest
            <i>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{yadav_asokan_yamamoto_patil_scott_2023, title={Expansion of the genetic toolbox for manipulation of the global crop pest <scp><i>Drosophila suzukii</i></scp>: 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} }
 @misc{patil_bhor_rhee_2020, title={Cell death in culture: Molecular mechanisms, detections, and inhibition strategies}, volume={91}, ISSN={["1876-794X"]}, DOI={10.1016/j.jiec.2020.08.009}, abstractNote={Mammalian cell cultures are widely used in the biopharmaceutical industry to produce monoclonal antibodies, vaccines, growth factors, etc. Cell death is an essential biological process for physiological growth and development, but it is a major problem in biopharmaceutical production in bio-industry. Cell death within bioreactor occurs due to various intracellular and extracellular stresses. These stresses negatively affect the culture longevity, overall product quality, and yield. Among all cell death types, apoptosis accounts for most of the cellular death in the bioreactor. The implementation and development of various strategies to prevent the cellular death are crucial for robust bioprocess development. Cell death during culture can be prevented or inhibited by supplementing media with specific chemicals, synthetic inhibitors, and genetic cell engineering approaches. In this review, we classified and described different types of cell death and their molecular mechanisms and summarized the cell death inhibition approaches implemented to inhibit cell death for various applications.}, journal={JOURNAL OF INDUSTRIAL AND ENGINEERING CHEMISTRY}, author={Patil, Anandrao Ashok and Bhor, Sachin Ashok and Rhee, Won Jong}, year={2020}, month={Nov}, pages={37–53} }