The compact Casπ (Cas12l) 'bracelet' provides a unique structural platform for DNA manipulation

被引：31
作者：
Sun, Ao [1 ,2 ,3 ]
Li, Cheng-Ping [1 ,2 ,3 ]
Chen, Zhihang [1 ,2 ,3 ]
Zhang, Shouyue [1 ,2 ,3 ]
Li, Dan-Yuan [1 ,2 ,3 ]
Yang, Yun [1 ,2 ,3 ]
Li, Long-Qi [1 ,2 ,3 ]
Zhao, Yuqian [1 ,2 ,3 ]
Wang, Kaichen [3 ]
Li, Zhaofu [3 ]
Liu, Jinxia [4 ,5 ]
Liu, Sitong [4 ,5 ]
Wang, Jia [1 ,2 ,3 ]
Liu, Jun-Jie Gogo [1 ,2 ,3 ]
机构：
[1] Tsinghua Univ, Beijing Adv Innovat Ctr Struct Biol, Beijing, Peoples R China
[2] Tsinghua Univ, Frontier Res Ctr Biol Struct, Sch Life Sci, Beijing, Peoples R China
[3] Tsinghua Univ, Tsinghua Peking Ctr Life Sci, Sch Life Sci, Beijing, Peoples R China
[4] Peking Univ, Dept Environm Engn, Beijing, Peoples R China
[5] Minist Educ China, Key Lab Water & Sediment Sci, Beijing, Peoples R China
来源：
CELL RESEARCH | 2023年 / 33卷 / 03期
基金：
国家重点研发计划; 中国国家自然科学基金;
关键词：
HUGE PHAGES; CRISPR; CLASSIFICATION; SYSTEMS; EVOLUTION;
D O I：
10.1038/s41422-022-00771-2
中图分类号：
Q2 [细胞生物学];
学科分类号：
071009 ; 090102 ;
摘要：
CRISPR-Cas modules serve as the adaptive nucleic acid immune systems for prokaryotes, and provide versatile tools for nucleic acid manipulation in various organisms. Here, we discovered a new miniature type V system, CRISPR-Cas pi (Cas12l) (similar to 860 aa), from the environmental metagenome. Complexed with a large guide RNA (similar to 170 nt) comprising the tracrRNA and crRNA, Cas pi (Cas12l) recognizes a unique 5 ' C-rich PAM for DNA cleavage under a broad range of biochemical conditions, and generates gene editing in mammalian cells. Cryo-EM study reveals a 'bracelet' architecture of Cas pi effector encircling the DNA target at 3.4 angstrom resolution, substantially different from the canonical 'two-lobe' architectures of Cas12 and Cas9 nucleases. The large guide RNA serves as a 'two-arm' scaffold for effector assembly. Our study expands the knowledge of DNA targeting mechanisms by CRISPR effectors, and offers an efficient but compact platform for DNA manipulation.
引用
收藏
页码：229 / 244
页数：16
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