Cas9-based genome editing in zebrafish.
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.
Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA; Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
Andrew P W
AP
Gonzales
Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA; Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA. Electronic address: jyeh1@mgh.harvard.edu.
Jing-Ruey Joanna
JR
Yeh
D000818:Animals; D064113:CRISPR-Cas Systems; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods; D016678:Genome; D054643:INDEL Mutation; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics; D015027:Zebrafish / Q000235:genetics*
D000094704:RNA, Guide, CRISPR-Cas Systems
CRISPR; Cas9; Chromosomal conversion; Chromosomal deletion; Gene-editing; Genome engineering; Homology-directed repair; Knock-in; Knock-out; Targeted integration; Targeted mutagenesis; Zebrafish