Cas9-based genome editing in zebrafish

• Published in: Methods in enzymology Vol. 546; p. 377
• Main Authors: Gonzales, Andrew P W, Yeh, Jing-Ruey Joanna
• Format: Journal Article
• Language: English
• Published: United States 2014
• Subjects: Animals; CRISPR-Cas Systems; Gene Targeting - methods; Genetic Engineering - methods; Genome; INDEL Mutation; RNA, Guide, CRISPR-Cas Systems - genetics; Zebrafish - genetics; CRISPR; Targeted mutagenesis; Genome engineering; Targeted integration; Chromosomal conversion; Chromosomal deletion; Zebrafish; Gene-editing; Cas9; Knock-in; Homology-directed repair; Knock-out
• ISSN: 1557-7988
• DOI: 10.1016/B978-0-12-801185-0.00018-0

Genome editing using the Cas9 endonuclease of Streptococcus pyogenes has demonstrated unprecedented efficacy and facility in a wide variety of biological systems. In zebrafish, specifically, studies have shown that Cas9 can be directed to user-defined genomic target sites via synthetic guide RNAs, enabling random or homology-directed sequence alterations, long-range chromosomal deletions, simultaneous disruption of multiple genes, and targeted integration of several kilobases of DNA. Altogether, these methods are opening new doors for the engineering of knock-outs, conditional alleles, tagged proteins, reporter lines, and disease models. In addition, the ease and high efficiency of generating Cas9-mediated gene knock-outs provides great promise for high-throughput functional genomics studies in zebrafish. In this chapter, we briefly review the origin of CRISPR/Cas technology and discuss current Cas9-based genome-editing applications in zebrafish with particular emphasis on their designs and implementations.