Open AccessCRISPR‐Cas9‐Guided Genome Engineering in C. elegans
Author(s) -
Kim HyunMin,
Colaiácovo Monica P.
Publication year - 2016
Publication title -
current protocols in molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.533
H-Index - 42
eISSN - 1934-3647
pISSN - 1934-3639
DOI - 10.1002/cpmb.7
Subject(s) - crispr , cas9 , genome editing , biology , genetics , homologous recombination , genome , computational biology , genome engineering , caenorhabditis elegans , dna , gene
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.