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Two high-fidelity variants: efSaCas9 and SaCas9-HF, which one is better?
- Source :
- Gene Therapy; August 2022, Vol. 29 Issue: 7-8 p458-463, 6p
- Publication Year :
- 2022
-
Abstract
- CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated endonuclease Cas9) nucleases have been widely applied for genome engineering. Staphylococcus aureus Cas9 (SaCas9) is compact, which can be packaged in AAV (adeno-associated virus) vector for in vivo gene editing. While, wild-type SaCas9 can induce unwanted off-target mutations and substantially limits the applications. So far, there are two reported SaCas9 variants with high-fidelity, including efSaCas9 from our previous study and SaCas9-HF. However, it remains unknown which one possessing the better fidelity and higher activity. Here, we performed a parallel comparison of efSaCas9 and SaCas9-HF in human cells through fluorescent reporter system and target deep sequencing, respectively. The results demonstrated that efSaCas9 possesses higher cleavage activity and fidelity than SaCas9-HF at the most endogenous sites in human cells. Collectively, our study provides insights for the rational selection of suitable SaCas9 for human genome editing.
Details
- Language :
- English
- ISSN :
- 09697128 and 14765462
- Volume :
- 29
- Issue :
- 7-8
- Database :
- Supplemental Index
- Journal :
- Gene Therapy
- Publication Type :
- Periodical
- Accession number :
- ejs58797837
- Full Text :
- https://doi.org/10.1038/s41434-022-00319-4