CRISPR‐Cas9‐Guided Genome Engineering in C. elegans. Issue 1 (1st July 2016)

• Record Type: Journal Article
• Authors: Kim, Hyun‐Min; Colaiácovo, Monica P.
• Editors: Ausubel, Frederick M.; Brent, Roger; Kingston, Robert E.; Moore, David D.; Seidman, J.G.; Smith, John A.; Struhl, Kevin
• Is Part Of: Current protocols in molecular biology. Volume 115:Issue 1(2016)
• Page Start: 31.7.1
• Page End: 31.7.18
• Publication Date: 2016-07-01
• Subjects: Cas9 -- C. elegans -- CRISPR -- CRISPR‐Cas -- genome editing -- genome engineering; Molecular biology -- Technique -- Periodicals; Molecular biology -- Laboratory manuals; Molecular Biology -- methods; Biologie moléculaire -- Technique; Biologie moléculaire -- Manuels de laboratoire; Molecular biology; Molecular biology -- Technique; Laboratory Manual; Electronic reference sources; Laboratory manuals; 572.8028
• DOI: 10.1002/cpmb.7
• Languages: English
• ISSNs: 1934-3639

Abstract

Abstract: 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.