RRC ID 49324
Author Haelterman NA, Jiang L, Li Y, Bayat V, Sandoval H, Ugur B, Tan KL, Zhang K, Bei D, Xiong B, Charng WL, Busby T, Jawaid A, David G, Jaiswal M, Venken KJ, Yamamoto S, Chen R, Bellen HJ.
Title Large-scale identification of chemically induced mutations in Drosophila melanogaster.
Journal Genome Res
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.
Volume 24(10)
Pages 1707-18
Published 2014-10-1
DOI 10.1101/gr.174615.114
PII gr.174615.114
PMID 25258387
PMC PMC4199363
MeSH Animals Chromosome Mapping / methods* Drosophila melanogaster / genetics* Ethyl Methanesulfonate Female Genes, Essential Genes, Insect Male Molecular Sequence Data Mutagenesis* Mutagens Polymorphism, Single Nucleotide Sequence Analysis, DNA / methods X Chromosome
IF 11.093
Times Cited 33