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

Science
Authors
Keywords
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
Journal
Science
Volume
353
Issue
6299
Pages
aaf5573
Date Published
2016 Aug 05
ISSN
1095-9203
URL
DOI
10.1126/science.aaf5573
PubMed ID
27256883
PubMed Central ID
PMC5127784
Links
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
Additional Materials