Cas9-based genome editing in zebrafish.

Abstract:

: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.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Gonzales AP,Yeh JR

doi

10.1016/B978-0-12-801185-0.00018-0

subject

Has Abstract

pub_date

2014-01-01 00:00:00

pages

377-413

eissn

0076-6879

issn

1557-7988

pii

B978-0-12-801185-0.00018-0

journal_volume

546

pub_type

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