CRISPR-Cas9-Guided Genome Engineering in C. elegans

Curr Protoc Mol Biol. 2016 Jul 1:115:31.7.1-31.7.18. doi: 10.1002/cpmb.7.


The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) system is successfully being used for efficient and targeted genome editing in various organisms, including the nematode C. elegans. Recent studies have developed various CRISPR-Cas9 approaches to enhance genome engineering via two major DNA double-strand break repair pathways: non-homologous end joining and homologous recombination. Here we describe a protocol for Cas9-mediated C. elegans genome editing together with single guide RNA (sgRNA) and repair template cloning, as well as injection methods required for delivering Cas9, sgRNAs, and repair template DNA into the C. elegans germline. © 2016 by John Wiley & Sons, Inc.

Keywords: C. elegans; CRISPR; CRISPR-Cas; Cas9; genome editing; genome engineering.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics
  • CRISPR-Cas Systems*
  • Caenorhabditis elegans / genetics*
  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Gene Editing / methods*
  • Genome
  • Homologous Recombination
  • RNA, Guide, CRISPR-Cas Systems / genetics


  • RNA, Guide, CRISPR-Cas Systems