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

Details

• Alternative Title:
  Science
• Personal Author:
  Abudayyeh, Omar O. ; Gootenberg, Jonathan S. ; Konermann, Silvana ; Joung, Julia ; Slaymaker, Ian M. ; Cox, David B.T. ; Shmakov, Sergey ; Makarova, Kira S. ; Semenova, Ekaterina ; Minakhin, Leonid ; Severinov, Konstantin ; Regev, Aviv ; Lander, Eric S. ; Koonin, Eugene V. ; Zhang, Feng
• Description:
  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.
• Subjects:
  Adaptive Immunity Bacterial Proteins Base Sequence Clustered Regularly Interspaced Short Palindromic Repeats CRISPR-Cas Systems Endoribonucleases Escherichia Coli Gene Knockdown Techniques Genetic Loci Leptotrichia Levivirus Molecular Sequence Data Protein Structure, Tertiary RNA, Guide, CRISPR-Cas Systems RNA Cleavage

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• Source:
  Science. 353(6299):aaf5573.
• Pubmed ID:
  27256883
• Pubmed Central ID:
  PMC5127784
• Document Type:
  Journal Article
• Funding:
  R01 MH110049/MH/NIMH NIH HHS/United States ; 5DP1-MH100706/DP/NCCDPHP CDC 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
• Volume:
  353
• Issue:
  6299
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:fc1cc85541cfa2de60b9870188d46d3213007b4af3bf6c6cefe565d8718962d7
• Download URL:
  https://stacks.cdc.gov/view/cdc/42943/cdc_42943_DS1.pdf
• File Type:
  [PDF - 1.78 MB ]