Journal
G3-GENES GENOMES GENETICS
Volume 6, Issue 8, Pages 2517-2521Publisher
GENETICS SOCIETY AMERICA
DOI: 10.1534/g3.116.031914
Keywords
CRISPR/Cas9; SaCas9; KKH SaCas9 variant; gene editing; zebrafish
Categories
Funding
- Science and Technology Program of Shenzhen [CXZZ20140903101756864, JCYJ20150924110425180, JCYJ201510301707 55264]
- China Postdoctoral Science Foundation
- International Postdoctoral Exchange Fellowship Program of China Postdoctoral Council [20130010]
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The type II CRISPR/Cas9 system has been used widely for genome editing in zebrafish. However, the requirement for the 59-NGG-39 protospacer-adjacent motif (PAM) of Cas9 from Streptococcus pyogenes (SpCas9) limits its targeting sequences. Here, we report that a Cas9 ortholog from Staphylococcus aureus (SaCas9), and its KKH variant, successfully induced targeted mutagenesis with high frequency in zebrafish. Confirming previous findings, the SpCas9 variant, VQR, can also induce targeted mutations in zebrafish. Bioinformatics analysis of these new Cas targets suggests that the number of available target sites in the zebrafish genome can be greatly expanded. Collectively, the expanded target repertoire of Cas9 in zebrafish should further facilitate the utility of this organism for genetic studies of vertebrate biology.
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