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

页码：229 / 244

页数：16

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