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Specific Gene Disruption in the Major Livestock Pests Cochliomyia hominivorax and Lucilia cuprina Using CRISPR/Cas9

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dc.contributor.author Paulo, Daniel F. en
dc.contributor.author Williamson, Megan E. en
dc.contributor.author Arp, Alex P. en
dc.contributor.author Li, Fang en
dc.contributor.author Sagel, Agustin en
dc.contributor.author Skoda, Steven R. en
dc.contributor.author Sanchez-Gallego, Joel en
dc.contributor.author Vasquez, Mario en
dc.contributor.author Quintero, Gladys en
dc.contributor.author Pérez, de Le en
dc.contributor.author Belikoff, Esther J. en
dc.contributor.author Azeredo-Espin, Ana en
dc.contributor.author McMillan, W. O. en
dc.contributor.author Concha, Carolina en
dc.contributor.author Scott, Maxwell J. en
dc.date.accessioned 2019-09-27T02:01:43Z
dc.date.available 2019-09-27T02:01:43Z
dc.date.issued 2019
dc.identifier.citation Paulo, Daniel F., Williamson, Megan E., Arp, Alex P., Li, Fang, Sagel, Agustin, Skoda, Steven R., Sanchez-Gallego, Joel, Vasquez, Mario, Quintero, Gladys, Pérez, de Le, Belikoff, Esther J., Azeredo-Espin, Ana, McMillan, W. O., Concha, Carolina, and Scott, Maxwell J. 2019. "Specific Gene Disruption in the Major Livestock Pests Cochliomyia hominivorax and Lucilia cuprina Using CRISPR/Cas9." <em>G3 (Bethesda, Md.)</em>. 9 (9):3045&ndash;3055. <a href="https://doi.org/10.1534/g3.119.400544">https://doi.org/10.1534/g3.119.400544</a> en
dc.identifier.issn 2160-1836
dc.identifier.uri https://hdl.handle.net/10088/98985
dc.description.abstract Cochliomyia hominivorax and Lucilia cuprina are major pests of livestock. Their larvae infest warm-blooded vertebrates and feed on host&#39;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 Chominivorax and Lcuprinayellow 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 Chominivoraxtransformer 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. en
dc.relation.ispartof G3 (Bethesda, Md.) en
dc.title Specific Gene Disruption in the Major Livestock Pests Cochliomyia hominivorax and Lucilia cuprina Using CRISPR/Cas9 en
dc.type Journal Article en
dc.identifier.srbnumber 152436
dc.identifier.doi 10.1534/g3.119.400544
rft.jtitle G3 (Bethesda, Md.)
rft.volume 9
rft.issue 9
rft.spage 3045
rft.epage 3055
dc.description.SIUnit STRI en
dc.description.SIUnit Peer-reviewed en
dc.citation.spage 3045
dc.citation.epage 3055

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