PUBLICATION
Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9
- Authors
- Feng, Y., Chen, C., Han, Y., Chen, Z., Lu, X., Liang, F., Li, S., Qin, W., Lin, S.
- ID
- ZDB-PUB-160619-5
- 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
-
- Gene Editing*
- Animals
- Zebrafish/embryology
- Zebrafish/genetics*
- Animals, Genetically Modified
- PubMed
- 27317783 Full text @ G3 (Bethesda)
Abstract
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.
Genes / Markers
Figure Gallery (1 images)
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping