Large-scale identification of chemically induced mutations in Drosophila melanogaster.

TitleLarge-scale identification of chemically induced mutations in Drosophila melanogaster.
Publication TypeJournal Article
Year of Publication2014
AuthorsHaelterman, NA, Jiang, L, Li, Y, Bayat, V, Sandoval, H, Ugur, B, Tan, KLi, Zhang, K, Bei, D, Xiong, B, Charng, W-L, Busby, T, Jawaid, A, David, G, Jaiswal, M, Venken, KJT, Yamamoto, S, Chen, R, Bellen, HJ
JournalGenome Res
Volume24
Issue10
Pagination1707-18
Date Published2014 Oct
ISSN1549-5469
KeywordsAnimals, Chromosome Mapping, Drosophila melanogaster, Ethyl Methanesulfonate, Female, Genes, Essential, Genes, Insect, Male, Molecular Sequence Data, Mutagenesis, Mutagens, Polymorphism, Single Nucleotide, Sequence Analysis, DNA, X Chromosome
Abstract

Forward genetic screens using chemical mutagens have been successful in defining the function of thousands of genes in eukaryotic model organisms. The main drawback of this strategy is the time-consuming identification of the molecular lesions causative of the phenotypes of interest. With whole-genome sequencing (WGS), it is now possible to sequence hundreds of strains, but determining which mutations are causative among thousands of polymorphisms remains challenging. We have sequenced 394 mutant strains, generated in a chemical mutagenesis screen, for essential genes on the Drosophila X chromosome and describe strategies to reduce the number of candidate mutations from an average of -3500 to 35 single-nucleotide variants per chromosome. By combining WGS with a rough mapping method based on large duplications, we were able to map 274 (-70%) mutations. We show that these mutations are causative, using small 80-kb duplications that rescue lethality. Hence, our findings demonstrate that combining rough mapping with WGS dramatically expands the toolkit necessary for assigning function to genes.

DOI10.1101/gr.174615.114
Alternate JournalGenome Res.
PubMed ID25258387
PubMed Central IDPMC4199363
Grant ListT32 HD055200 / HD / NICHD NIH HHS / United States
K12 GM084897 / GM / NIGMS NIH HHS / United States
5R01GM067858 / GM / NIGMS NIH HHS / United States
R21 HG006726 / HG / NHGRI NIH HHS / United States
RC4 GM096355 / GM / NIGMS NIH HHS / United States
U54 HD083092 / HD / NICHD NIH HHS / United States
5K12GM084897 / GM / NIGMS NIH HHS / United States
1RC4GM096355-01 / GM / NIGMS NIH HHS / United States
/ / Howard Hughes Medical Institute / United States
R01 GM067858 / GM / NIGMS NIH HHS / United States
5T32-HD055200 / HD / NICHD NIH HHS / United States
1R21HG006726 / HG / NHGRI NIH HHS / United States