Structural basis of negative regulation of CRISPR-Cas7-11 by TPR-CHAT

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外文摘要：CRISPR-Cas7-11 is a Type Ⅲ-E CRISPR-associated nuclease that functions as a potent RNA editing tool.Tetratrico-peptide repeat fused with Cas/HEF1-associated signal transducer(TPR-CHAT)acts as a regulatory protein that interacts with CRISPR RNA(crRNA)-bound Cas7-11 to form a CRISPR-guided caspase complex(Craspase).However,the precise modulation of Cas7-11's nuclease activity by TPR-CHAT to enhance its utility requires further study.Here,we report cryo-electron microscopy(cryo-EM)structures of Desulfonema ishimotonii(Di)Cas7-11-crRNA,complexed with or without the full length or the N-terminus of TPR-CHAT.These structures unveil the molecular features of the Craspase complex.Structural analysis,combined with in vitro nuclease assay and electrophoretic mobility shift assay,reveals that DiTPR-CHAT negatively regulates the activity of DiCas7-11 by preventing target RNA from binding through the N-terminal 65 amino acids of DiTPR-CHAT(DiTPR-CHATNTD).Our work demonstrates that DiTPR-CHATNTD can function as a small unit of DiCas7-11 regulator,potentially enabling safe applications to prevent overcutting and off-target effects of the CRISPR-Cas7-11 system.

作者：

Tian Hong、Qinghua Luo、Haiyun Ma、Xin Wang、Xinqiong Li、Chongrong Shen、Jie Pang、Yan Wang、Yuejia Chen、Changbin Zhang、Zhaoming Su、Haohao Dong、Xiaodi Tang

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作者单位：

Department of Laboratory Medicine,State Key Laboratory of Biotherapy,National Clinical Research Center for Geriatrics,West China Hospital,Sichuan University,Chengdu,China

Frontiers Medical Center,Tianfu Jincheng Laboratory,West China Hospital,Sichuan University,Chengdu,China

基金：

National Key Research and Development Program of ChinaNational Key Research and Development Program of ChinaNational Key Research and Development Program of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of China1.3.5 Project for Disciplines Excellence of West China Hospital,Sichuan UniversityTianjin Synthetic Biotechnology Innovation Capacity Improvement Action

项目编号：

2021YFA13019002021YFA13012032022YFC23037003190003932170029819719743222204032070049ZYYC20021TSBICIP-KJGG-008

出版年：

2024

DOI：

10.1038/s41392-024-01821-4

信号转导与靶向治疗(英文)

CSTPCD

ISSN：

年,卷(期)：2024.9(6)