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dc.contributor.authorHighfill, Chad Allen
dc.contributor.authorTran, Jonathan H.
dc.contributor.authorNguyen, Samantha K. T.
dc.contributor.authorMoldenhauer, Taylor R.
dc.contributor.authorWang, Xiaofei
dc.contributor.authorMacdonald, Stuart J.
dc.date.accessioned2018-12-13T21:27:57Z
dc.date.available2018-12-13T21:27:57Z
dc.date.issued2017-07-10
dc.identifier.citationNaturally Segregating Variation at Ugt86Dd Contributes to Nicotine Resistance in Drosophila melanogaster Chad A. Highfill, Jonathan H. Tran, Samantha K. T. Nguyen, Taylor R. Moldenhauer, Xiaofei Wang and View ORCID ProfileStuart J. Macdonald Genetics September 1, 2017 vol. 207 no. 1 311-325; https://doi.org/10.1534/genetics.117.300058en_US
dc.identifier.urihttp://hdl.handle.net/1808/27503
dc.description.abstractIdentifying the sequence polymorphisms underlying complex trait variation is a key goal of genetics research, since knowing the precise causative molecular events allows insight into the pathways governing trait variation. Genetic analysis of complex traits in model systems regularly starts by constructing QTL maps, but generally fails to identify causative sequence polymorphisms. Previously we mapped a series of QTL contributing to resistance to nicotine in a Drosophila melanogaster multiparental mapping resource and here use a battery of functional tests to resolve QTL to the molecular level. One large-effect QTL resided over a cluster of UDP-glucuronosyltransferases, and quantitative complementation tests using deficiencies eliminating subsets of these detoxification genes revealed allelic variation impacting resistance. RNAseq showed that Ugt86Dd had significantly higher expression in genotypes that are more resistant to nicotine, and anterior midgut-specific RNA interference (RNAi) of this gene reduced resistance. We discovered a segregating 22-bp frameshift deletion in Ugt86Dd, and accounting for the InDel during mapping largely eliminates the QTL, implying the event explains the bulk of the effect of the mapped locus. CRISPR/Cas9 editing of a relatively resistant genotype to generate lesions in Ugt86Dd that recapitulate the naturally occurring putative loss-of-function allele, leads to a large reduction in resistance. Despite this major effect of the deletion, the allele appears to be very rare in wild-caught populations and likely explains only a small fraction of the natural variation for the trait. Nonetheless, this putatively causative coding InDel can be a launchpad for future mechanistic exploration of xenobiotic detoxification.en_US
dc.publisherASTM Internationalen_US
dc.rightsCopyright © 2017 Highfill. Available freely online through the author-supported open access option.This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.en_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_US
dc.subjectXenobioticsen_US
dc.subjectCRISPRen_US
dc.subjectFunctional validationen_US
dc.subjectQTL mappingen_US
dc.subjectRNAien_US
dc.titleNaturally Segregating Variation at Ugt86Dd Contributes to Nicotine Resistance in Drosophila melanogasteren_US
dc.typeArticleen_US
kusw.kudepartmentMolecular Biosciencesen_US
dc.identifier.doiGenetics Society of Americaen_US
kusw.oaversionScholarly/refereed, publisher versionen_US
kusw.oapolicyThis item meets KU Open Access policy criteria.en_US
dc.rights.accessrightsopenAccessen_US


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Copyright © 2017 Highfill. Available freely online through the author-supported open access option.This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Except where otherwise noted, this item's license is described as: Copyright © 2017 Highfill. Available freely online through the author-supported open access option.This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.