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A naturally-occurring 22-bp coding deletion in Ugt86Dd reduces nicotine resistance in Drosophila melanogaster

Macdonald, Stuart J.
; 
Highfill, Chad A.
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Abstract
Objective Segregating genetic variants contribute to the response to toxic, xenobiotic compounds, and identifying these causative sites can help describe the mechanisms underlying metabolism of toxic compounds. In previous work we implicated the detoxification gene Ugt86Dd in the genetic control of larval nicotine resistance in Drosophila melanogaster. Furthermore, we suggested that a naturally-occurring 22-bp deletion that leads to a stop codon in exon 2 of the gene markedly reduces resistance. Here we use homology directed CRISPR/Cas9 gene editing to specifically test this hypothesis. Results We edited chromosome three from an inbred strain named A4 which carries the insertion allele at Ugt86Dd, successfully generated four alleles carrying the 22-bp Ugt86Dd deletion, and substituted edited chromosomes back into the A4 background. The original A4 strain, and an un-edited control strain in the same A4 background, show no significant difference in egg-to-adult or larva-to-adult viability on either control media or nicotine-supplemented media, and only slightly delayed development in nicotine media. However, strains carrying the 22-bp deletion showed reduced viability in nicotine conditions, and significantly longer development. Our data strongly suggest that the naturally-occurring 22-bp insertion/deletion event in Ugt86Dd directly impacts variation in nicotine resistance in D. melanogaster.
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This work is licensed under a Creative Commons Attribution 4.0 International License.
Date
2020-03-30
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BMC
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Molecular Biosciences Scholarly Works
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Keywords
Ugt86Dd, Ugt35C1, Drosophila, Nicotine, CRISPR, QTL, DSPR
Citation
Macdonald, S. J., & Highfill, C. A. (2020). A naturally-occurring 22-bp coding deletion in Ugt86Dd reduces nicotine resistance in Drosophila melanogaster. BMC research notes, 13(1), 188. https://doi.org/10.1186/s13104-020-05035-z
URI
https://hdl.handle.net/1808/30538
DOI
10.1186/s13104-020-05035-z
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