RNA targeting unleashes indiscriminate nuclease activity of CRISPR-Cas12a2.
RNA targeting unleashes indiscriminate nuclease activity of CRISPR-Cas12a2.
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DOI：
10.1038/s41586-022-05560-w
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发表时间：
2023-01
期刊：
影响因子：
64.8
通讯作者：
Taylor, David W.
中科院分区：
文献类型：
--
作者：
Bravo, Jack P. K.;Hallmark, Thomson;Naegle, Bronson;Beisel, Chase L.;Jackson, Ryan N.;Taylor, David W.
Cas12a2是一种crispr相关核酸酶，在识别互补RNA靶标后，对单链RNA、单链DNA和双链DNA进行RNA引导的序列非特异性降解，最终导致流产感染。本文报道了Cas12a2在二元、三元和四元配合物中的结构，揭示了其完整的激活途径。我们的结构揭示了Cas12a2在与同源RNA靶标结合之前是自动抑制的，这使得RuvC活性位点暴露在一个大的带正电的裂缝中。双链DNA底物通过双扭曲和局部熔化被捕获，由对双链DNA降解和体内免疫系统功能至关重要的“芳香钳”残基对稳定。我们的工作为这种流产感染机制提供了结构基础，以实现群体水平的免疫，这可以用来创造合理的突变体，降解一系列附带底物。Cas12a2是一种非特异性切割ssRNA、ssDNA和dsDNA的crispr相关核酸酶，其结构分析揭示了一个完整的激活途径，参与了流产感染系统保护细胞免受入侵的过程。
Cas12a2 is a CRISPR-associated nuclease that performs RNA-guided, sequence-nonspecific degradation of single-stranded RNA, single-stranded DNA and double-stranded DNA following recognition of a complementary RNA target, culminating in abortive infection. Here we report structures of Cas12a2 in binary, ternary and quaternary complexes to reveal a complete activation pathway. Our structures reveal that Cas12a2 is autoinhibited until binding a cognate RNA target, which exposes the RuvC active site within a large, positively charged cleft. Double-stranded DNA substrates are captured through duplex distortion and local melting, stabilized by pairs of ‘aromatic clamp’ residues that are crucial for double-stranded DNA degradation and in vivo immune system function. Our work provides a structural basis for this mechanism of abortive infection to achieve population-level immunity, which can be leveraged to create rational mutants that degrade a spectrum of collateral substrates. Structural analysis of Cas12a2, a CRISPR-associated nuclease that nonspecifically cleaves ssRNA, ssDNA and dsDNA, reveals a complete activation pathway involved in the abortive infection system protecting cells against invasion.
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影响因子：
64.8
作者：
Dmytrenko, Oleg;Neumann, Gina C.;Hallmark, Thomson;Keiser, Dylan J.;Crowley, Valerie M.;Vialetto, Elena;Mougiakos, Ioannis;Wandera, Katharina G.;Domgaard, Hannah;Weber, Johannes;Gaudin, Thomas;Metcalf, Josie;Gray, Benjamin N.;Begemann, Matthew B.;Jackson, Ryan N.;Beisel, Chase L.
通讯作者：
Beisel, Chase L.
DOI：
10.1126/science.aaf5573
发表时间：
2016-08-05
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Abudayyeh OO;Gootenberg JS;Konermann S;Joung J;Slaymaker IM;Cox DB;Shmakov S;Makarova KS;Semenova E;Minakhin L;Severinov K;Regev A;Lander ES;Koonin EV;Zhang F
通讯作者：
Zhang F
影响因子：
64.8
作者：
Jumper J;Evans R;Pritzel A;Green T;Figurnov M;Ronneberger O;Tunyasuvunakool K;Bates R;Žídek A;Potapenko A;Bridgland A;Meyer C;Kohl SAA;Ballard AJ;Cowie A;Romera-Paredes B;Nikolov S;Jain R;Adler J;Back T;Petersen S;Reiman D;Clancy E;Zielinski M;Steinegger M;Pacholska M;Berghammer T;Bodenstein S;Silver D;Vinyals O;Senior AW;Kavukcuoglu K;Kohli P;Hassabis D
通讯作者：
Hassabis D
CRISPR-C2C2的两种不同的RNase活性启用了指导RNA处理和RNA检测。
影响因子：
64.8
作者：
East-Seletsky A;O'Connell MR;Knight SC;Burstein D;Cate JH;Tjian R;Doudna JA
通讯作者：
Doudna JA
影响因子：
14.9
作者：
Harris CJ;Molnar A;Müller SY;Baulcombe DC
通讯作者：
Baulcombe DC