Caenorhabditis elegans mutant allele identification by whole-genome sequencing

Abstract

Identification of the molecular lesion in Caenorhabditis elegans mutants isolated through forward genetic screens usually involves time-consuming genetic mapping. We used Illumina deep sequencing technology to sequence a complete, mutant C. elegans genome and thus pinpointed a single-nucleotide mutation in the genome that affects a neuronal cell fate decision. This constitutes a proof-of-principle for using whole-genome sequencing to analyze C. elegans mutants.

Figure 1

Variants found in whole-genome sequence analysis. ot177 is shorthand for lsy-12(ot177). The asterisk denotes 6 variants clustered within 100 bp of a single intron (supplementary Table 1a), which upon amplification and Sanger-sequencing we found to map at least in part onto a different chromosome.

Figure 2

Physical location of the lsy-12 locus and its lesions. lsy-12(ot177) was mapped between two SNP markers, pkP5112 and pkP5064 (http://www.wormbase.org/), which define a 4-Mb interval. Validated exonic variants in this interval are marked with arrowheads. Numbers indicate base position on chromosome V. The gene R07B5.9, identified as lsy-12, is shown below and the positions of the previously identified lsy-12 alleles are marked with black arrows.