Engineering the Caenorhabditis elegans genome with CRISPR/Cas9

Methods. 2014 Aug 1;68(3):381-8. doi: 10.1016/j.ymeth.2014.03.024. Epub 2014 Mar 28.


The development in early 2013 of CRISPR/Cas9-based genome engineering promises to dramatically advance our ability to alter the genomes of model systems at will. A single, easily produced targeting RNA guides the Cas9 endonuclease to a specific DNA sequence where it creates a double strand break. Imprecise repair of the break can yield mutations, while homologous recombination with a repair template can be used to effect specific changes to the genome. The tremendous potential of this system led several groups to independently adapt it for use in Caenorhabditiselegans, where it was successfully used to generate mutations and to create tailored genome changes through homologous recombination. Here, we review the different approaches taken to adapt CRISPR/Cas9 for C. elegans, and provide practical guidelines for CRISPR/Cas9-based genome engineering.

Keywords: CRISPR; Caenorhabditis elegans; Cas9; Double strand break; Genome engineering; Homologous recombination.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caenorhabditis elegans / genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • DNA Breaks, Double-Stranded*
  • Gene Targeting
  • Genetic Engineering / methods*
  • Genome
  • Homologous Recombination / genetics*
  • Mutagenesis