ZFIN ID: ZDB-PUB-160619-5
Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9
Feng, Y., Chen, C., Han, Y., Chen, Z., Lu, X., Liang, F., Li, S., Qin, W., Lin, S.
Date: 2016
Source: G3 (Bethesda)   6(8): 2517-21 (Journal)
Registered Authors: Lin, Shuo
Keywords: CRISPR/Cas9, KKH SaCas9 variant, SaCas9, gene editing, zebrafish
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Bacterial Proteins/genetics
  • CRISPR-Cas Systems*
  • Embryo, Nonmammalian
  • Gene Editing*
  • Genome*
  • Staphylococcus aureus/genetics
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed: 27317783 Full text @ G3 (Bethesda)
The type II CRISPR/Cas9 system has been widely used for genome editing in zebrafish. However, the requirement for 5'-NGG-3' protospacer-adjacent motif (PAM) of Cas9 from Streptococcus pyogenes (SpCas9) limits its targeting sequences. Here, we report that a Cas9 orthologue from Staphylococcus aureus (SaCas9) and its KKH variant successfully induced targeted mutagenesis with high frequency in zebrafish. Confirming the previous finding, 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 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.