C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector.

C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector.
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DOI:
10.1126/science.aaf5573
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发表时间:
2016-08-05
期刊:
Science (New York, N.Y.)
影响因子:
--
通讯作者:
Zhang F
Zhang F
中科院分区:
其他
文献类型:
--
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

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CRISPR-Cas适应性免疫系统通过DNA或RNA-DNA干扰保护微生物免受外来遗传因素的影响。我们对Class2Type VI-A CRISPR-Cas效应器C2c2进行了表征,并展示了其RNA引导的核糖核酸酶功能。来自细毛菌沙希菌的C2c2对RNA噬菌体提供干扰。体外生化分析表明,C2c2是由单个crRNA引导的,可以编程切割携带互补蛋白的单链RNA靶标。在细菌中,C2c2可以被编程为破坏特定的mRNA。切割是由两个保守的HEPN结构域上的催化残基介导的,在这些结构域中突变会产生催化失活的RNA结合蛋白。这些结果拓宽了我们对CRISPR-CAS系统的理解,并表明C2c2可用于开发新的RNA靶向工具。
The CRISPR-Cas adaptive immune system defends microbes against foreign genetic elements via DNA or RNA-DNA interference. We characterize the Class 2 type VI-A CRISPR-Cas effector C2c2 and demonstrate its RNA-guided RNase function. C2c2 from the bacterium Leptotrichia shahii provides interference against RNA phage. In vitro biochemical analysis show that C2c2 is guided by a single crRNA and can be programmed to cleave ssRNA targets carrying complementary protospacers. In bacteria, C2c2 can be programmed to knock down specific mRNAs. Cleavage is mediated by catalytic residues in the two conserved HEPN domains, mutations in which generate catalytically inactive RNA-binding proteins. These results broaden our understanding of CRISPR-Cas systems and suggest that C2c2 can be used to develop new RNA-targeting tools.
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