Cas12a2 elicits abortive infection through RNA-triggered destruction of dsDNA.
Cas12a2 elicits abortive infection through RNA-triggered destruction of dsDNA.
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DOI：
10.1038/s41586-022-05559-3
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发表时间：
2023-01
期刊：
影响因子：
64.8
通讯作者：
Beisel, Chase L.
中科院分区：
文献类型：
--
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.
细菌的失败感染系统通过在入侵者复制之前关闭或杀死受感染的细胞来限制外来入侵者的传播。几种靶向RNA的CRISPR-Cas系统（即III型和VI型）通过激活不加选择的核酸酶引起流产感染表型。然而，尚未观察到CRISPR介导的利用RNA引导的单效应物核酸酶的不加选择的DNA酶活性的流产机制。在这里，我们报告了V型单效应核酸酶Cas 12 a2的RNA靶向通过双链DNA（dsDNA）的非特异性切割驱动流产感染。在识别具有激活的前间隔区侧翼序列的RNA靶标后，Cas 12 a2有效地降解单链RNA（ssRNA）、单链DNA（ssDNA）和dsDNA。在细胞内，Cas 12 a2的激活诱导SOS DNA损伤反应并损害生长，阻止入侵者的传播。最后，我们利用Cas 12 a2的间接活性进行直接RNA检测，证明Cas 12 a2可以重新用作RNA引导的RNA靶向工具。这些发现扩展了CRISPR-Cas系统已知的防御能力，并为CRISPR技术创造了更多的机会。硫卷叶菌V型单效应核酸酶SuCas 12 a2的RNA靶向作用通过双链DNA的非特异性切割驱动流产感染-在通过激活的前间隔区侧翼序列识别RNA靶点后，SuCas 12 a2有效降解ssRNA、ssDNA和dsDNA。
Bacterial abortive-infection systems limit the spread of foreign invaders by shutting down or killing infected cells before the invaders can replicate. Several RNA-targeting CRISPR–Cas systems (that is, types III and VI) cause abortive-infection phenotypes by activating indiscriminate nucleases. However, a CRISPR-mediated abortive mechanism that leverages indiscriminate DNase activity of an RNA-guided single-effector nuclease has yet to be observed. Here we report that RNA targeting by the type V single-effector nuclease Cas12a2 drives abortive infection through non-specific cleavage of double-stranded DNA (dsDNA). After recognizing an RNA target with an activating protospacer-flanking sequence, Cas12a2 efficiently degrades single-stranded RNA (ssRNA), single-stranded DNA (ssDNA) and dsDNA. Within cells, the activation of Cas12a2 induces an SOS DNA-damage response and impairs growth, preventing the dissemination of the invader. Finally, we harnessed the collateral activity of Cas12a2 for direct RNA detection, demonstrating that Cas12a2 can be repurposed as an RNA-guided RNA-targeting tool. These findings expand the known defensive abilities of CRISPR–Cas systems and create additional opportunities for CRISPR technologies. RNA targeting by the Sulfuricurvum type V single-effector nuclease SuCas12a2 drives abortive infection through non-specific cleavage of double-stranded DNA—after recognition of an RNA target through an activating protospacer-flanking sequence, SuCas12a2 efficiently degrades ssRNA, ssDNA and dsDNA.
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影响因子：
16.6
作者：
Collias D;Beisel CL
通讯作者：
Beisel CL
影响因子：
14.9
作者：
Han W;Li Y;Deng L;Feng M;Peng W;Hallstrøm S;Zhang J;Peng N;Liang YX;White MF;She Q
通讯作者：
She Q
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
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：
14.8
作者：
Kelley LA;Mezulis S;Yates CM;Wass MN;Sternberg MJ
通讯作者：
Sternberg MJ