C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector.
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Abstract | The clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated genes (Cas) adaptive immune system defends microbes against foreign genetic elements via DNA or RNA-DNA interference. We characterize the class 2 type VI CRISPR-Cas effector C2c2 and demonstrate its RNA-guided ribonuclease function. C2c2 from the bacterium Leptotrichia shahii provides interference against RNA phage. In vitro biochemical analysis shows that C2c2 is guided by a single CRISPR RNA and can be programmed to cleave single-stranded RNA 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 Higher Eukaryotes and Prokaryotes Nucleotide-binding (HEPN) domains, mutations of 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. |
Year of Publication | 2016
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Journal | Science
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Volume | 353
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Issue | 6299
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Pages | aaf5573
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Date Published | 2016 Aug 05
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ISSN | 1095-9203
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URL | |
DOI | 10.1126/science.aaf5573
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PubMed ID | 27256883
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PubMed Central ID | PMC5127784
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Grant list | R01 MH110049 / MH / NIMH NIH HHS / United States
5DP1-MH100706 / DP / NCCDPHP CDC HHS / United States
P30 CA014051 / CA / NCI NIH HHS / United States
T32 GM007753 / GM / NIGMS NIH HHS / United States
Intramural NIH HHS / United States
1R01-MH110049 / MH / NIMH NIH HHS / United States
DP1 MH100706 / MH / NIMH NIH HHS / United States
T32GM007753 / GM / NIGMS NIH HHS / United States
GM10407 / GM / NIGMS NIH HHS / United States
RM1 HG006193 / HG / NHGRI NIH HHS / United States
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