Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination

Nucleic Acids Res. 2013 Nov;41(20):e193. doi: 10.1093/nar/gkt805. Epub 2013 Sep 5.

Abstract

Cas9 is an RNA-guided double-stranded DNA nuclease that participates in clustered regularly interspaced short palindromic repeats (CRISPR)-mediated adaptive immunity in prokaryotes. CRISPR-Cas9 has recently been used to generate insertion and deletion mutations in Caenorhabditis elegans, but not to create tailored changes (knock-ins). We show that the CRISPR-CRISPR-associated (Cas) system can be adapted for efficient and precise editing of the C. elegans genome. The targeted double-strand breaks generated by CRISPR are substrates for transgene-instructed gene conversion. This allows customized changes in the C. elegans genome by homologous recombination: sequences contained in the repair template (the transgene) are copied by gene conversion into the genome. The possibility to edit the C. elegans genome at selected locations will facilitate the systematic study of gene function in this widely used model organism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems
  • Caenorhabditis elegans / genetics*
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Deoxyribonucleases / metabolism
  • Gene Conversion
  • Gene Targeting / methods*
  • Genetic Engineering / methods
  • Genome
  • Mutagenesis
  • RNA, Guide / metabolism
  • Recombinational DNA Repair*
  • Transgenes

Substances

  • RNA, Guide
  • Deoxyribonucleases