| Autor: |
Yangcan Chen, Yanping Hu, Xinge Wang, Shengqiu Luo, Ning Yang, Yi Chen, Zhikun Li, Qi Zhou, Wei Li |
| Jazyk: |
angličtina |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
The Innovation, Vol 3, Iss 4, Pp 100264- (2022) |
| Druh dokumentu: |
article |
| ISSN: |
2666-6758 |
| DOI: |
10.1016/j.xinn.2022.100264 |
| Popis: |
The naturally occurring prokaryotic CRISPR-Cas systems provide valuable resources for the development of new genome-editing tools. However, the majority of prokaryotic Cas nucleases exhibit poor editing efficiency in mammalian cells, which significantly limits their utility. Here, we have developed a method termed Improving Editing Activity by Synergistic Engineering (MIDAS). This method exerts a synergistic effect to improve mammalian genome-editing efficiency of a wide range of CRISPR-Cas systems by enhancing the interactions between Cas nuclease with the protospacer adjacent motif (PAM) and the single-stranded DNA (ssDNA) substrate in the catalytic pocket simultaneously. MIDAS robustly and significantly increased the gene-editing efficiency of Cas12i, Cas12b, and CasX in human cells. Notably, a Cas12i variant, Cas12iMax, exhibited robust activity with a very broad PAM range (NTNN, NNTN, NAAN, and NCAN) and higher efficiency than the current widely used Cas nucleases. A high-fidelity version of Cas12iMax (Cas12iHiFi) has been further engineered to minimize off-target effects. Our work provides an expandable and efficacious method for engineering Cas nucleases for robust mammalian genome editing. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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