清脆的
基因组编辑
Cas9
核酸酶
核糖核蛋白
计算生物学
基因组工程
质粒
生物
基因组
DNA
引导RNA
核糖核酸
遗传学
基因
作者
Zhaowei Wu,Yifei Zhang,Haopeng Yu,Deng Pan,Yujue Wang,Yannan Wang,Fan Li,Chang Liu,Hao Nan,Weizhong Chen,Quanjiang Ji
标识
DOI:10.1038/s41589-021-00868-6
摘要
The RNA-guided CRISPR-associated (Cas) nucleases are versatile tools for genome editing in various organisms. The large sizes of the commonly used Cas9 and Cas12a nucleases restrict their flexibility in therapeutic applications that use the cargo-size-limited adeno-associated virus delivery vehicle. More compact systems would thus offer more therapeutic options and functionality for this field. Here, we report a miniature class 2 type V-F CRISPR-Cas genome-editing system from Acidibacillus sulfuroxidans (AsCas12f1, 422 amino acids). AsCas12f1 is an RNA-guided endonuclease that recognizes 5′ T-rich protospacer adjacent motifs and creates staggered double-stranded breaks to target DNA. We show that AsCas12f1 functions as an effective genome-editing tool in both bacteria and human cells using various delivery methods, including plasmid, ribonucleoprotein and adeno-associated virus. The small size of AsCas12f1 offers advantages for cellular delivery, and characterizations of AsCas12f1 may facilitate engineering more compact genome-manipulation technologies.
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