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

CRISPR-Cas9-Guided Genome Engineering in C. elegans.

Author information

1
Department of Genetics, Harvard Medical School, Boston, Massachusetts.

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.

KEYWORDS:

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

PMID:
27366893
PMCID:
PMC4932911
DOI:
10.1002/cpmb.7
[Indexed for MEDLINE]
Free PMC Article

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