Efficient target-selected mutagenesis in Caenorhabditis elegans: toward a knockout for every gene.

ABSTRACT: Reverse genetic or gene-driven knockout approaches have contributed significantly to the success of model organisms for fundamental and biomedical research. Although various technologies are available for C. elegans, none of them scale very well for genome-wide application. To address this, we implemented a target-selected knockout approach that is based on random chemical mutagenesis and detection of single nucleotide mutations in genes of interest using high-throughput resequencing. A clonal library of 6144 EMS-mutagenized worms was established and screened, resulting in the identification of 1044 induced mutations in 109 Mbp, which translates into an average spacing between exonic mutations in the library of only 17 bp. We covered 25% of the open reading frames of 32 genes and identifie

SUBMITTER: Cuppen E 

PROVIDER: S-EPMC1855173 | biostudies-literature | 2007 May

REPOSITORIES: biostudies-literature