Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9.
The type II CRISPR/Cas9 system has been used widely for genome editing in zebrafish. However, the requirement for the 5'-NGG-3' 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.
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China.
Yan
Y
Feng
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China.
Cheng
C
Chen
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China.
Yuxiang
Y
Han
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, California 90095.
Zelin
Z
Chen
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China.
Xiaochan
X
Lu
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China.
Fang
F
Liang
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China.
Song
S
Li
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China qinwei@pkusz.edu.cn shuolin@ucla.edu.
Wei
W
Qin
Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, California 90095 qinwei@pkusz.edu.cn shuolin@ucla.edu.
Shuo
S
Lin
D000818:Animals; D030801:Animals, Genetically Modified; D001426:Bacterial Proteins / Q000235:genetics; D064113:CRISPR-Cas Systems; D004625:Embryo, Nonmammalian; D000072669:Gene Editing; D016678:Genome; D013211:Staphylococcus aureus / Q000235:genetics; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*
D001426:Bacterial Proteins
CRISPR/Cas9; KKH SaCas9 variant; SaCas9; gene editing; zebrafish