清脆的
生物
核糖核酸
DNA
基因组编辑
效应器
劈开
计算生物学
核酸
基因组
Cas9
引导RNA
基因组工程
遗传学
细胞生物学
基因
作者
Satoru N. Takeda,Ryoya Nakagawa,Sae Okazaki,Hisato Hirano,Kan Kobayashi,Tsukasa Kusakizako,Tomohiro Nishizawa,Keitaro Yamashita,Hiroshi Nishimasu,Osamu Nureki
出处
期刊:Molecular Cell
[Elsevier]
日期:2020-12-17
卷期号:81 (3): 558-570.e3
被引量:128
标识
DOI:10.1016/j.molcel.2020.11.035
摘要
RNA-guided DNA endonucleases derived from CRISPR-Cas adaptive immune systems are widely used as powerful genome-engineering tools. Among the diverse CRISPR-Cas nucleases, the type V-F Cas12f (also known as Cas14) proteins are exceptionally compact and associate with a guide RNA to cleave single- and double-stranded DNA targets. Here, we report the cryo-electron microscopy structure of Cas12f1 (also known as Cas14a) in complex with a guide RNA and its target DNA. Unexpectedly, the structure revealed that two Cas12f1 molecules assemble with the single guide RNA to recognize the double-stranded DNA target. Each Cas12f1 protomer adopts a different conformation and plays distinct roles in nucleic acid recognition and DNA cleavage, thereby explaining how the miniature Cas12f1 enzyme achieves RNA-guided DNA cleavage as an “asymmetric homodimer.” Our findings augment the mechanistic understanding of diverse CRISPR-Cas nucleases and provide a framework for the development of compact genome-engineering tools critical for therapeutic genome editing.
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