Two High-fidelity Variants: EfSaCas9 and SaCas9-HF, Which One is Better?

Overview

Journal Gene Ther

Date 2022 Jan 31

PMID 35095097

Authors

Jineng Lv

Haitao Xi

Xiujuan Lv

Yue Zhou

Jiahua Wang

Haoran Chen

Tong Yan

Jiang Jin

Junzhao Zhao

Feng Gu

Zongming Song

Affiliations

Soon will be listed here.

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.

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