Discovery and functional characterization of diverse Class 2 CRISPR-Cas systems

Details

• Alternative Title:
  Mol Cell
• Personal Author:
  Shmakov, Sergey ; Abudayyeh, Omar O. ; Makarova, Kira S. ; Wolf, Yuri I ; Gootenberg, Jonathan S. ; Semenova, Ekaterina ; Minakhin, Leonid ; Joung, Julia ; Konermann, Silvana ; Severinov, Konstantin ; Zhang, Feng ; Koonin, Eugene V.
• Description:
  Microbial CRISPR-Cas systems are divided into Class 1, with multisubunit effector complexes, and Class 2, with single protein effectors. Currently, only two Class 2 effectors, Cas9 and Cpf1, are known. We describe here three distinct Class 2 CRISPR-Cas systems. The effectors of two of the identified systems, C2c1 and C2c3, contain RuvC-like endonuclease domains distantly related to Cpf1. The third system, C2c2, contains an effector with two predicted HEPN RNase domains. Whereas production of mature CRISPR RNA (crRNA) by C2c1 depends on tracrRNA, C2c2 crRNA maturation is tracrRNA independent. We found that C2c1 systems can mediate DNA interference in a 5'-PAM-dependent fashion analogous to Cpf1. However, unlike Cpf1, which is a single-RNA-guided nuclease, C2c1 depends on both crRNA and tracrRNA for DNA cleavage. Finally, comparative analysis indicates that Class 2 CRISPR-Cas systems evolved on multiple occasions through recombination of Class 1 adaptation modules with effector proteins acquired from distinct mobile elements.
• Subjects:
  Article Bacteria Bacterial Proteins Cas9 Computational Discovery Pipeline Cpf1 CRISPR-Cas Adaptive Immunity CRISPR-Cas Systems CrRNA Evolution, Molecular HEPN Domain PAM Protein Structure, Tertiary Recombination, Genetic Ribonucleases RNA, Bacterial RNA-seq RuvC-like Endonuclease TracrRNA

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• Source:
  Mol Cell. 60(3):385-397.
• Pubmed ID:
  26593719
• Pubmed Central ID:
  PMC4660269
• Document Type:
  Journal Article
• Funding:
  R01 MH110049/MH/NIMH NIH HHS/United States ; 5DP1-MH100706/DP/NCCDPHP CDC HHS/United States ; R01 GM104071/GM/NIGMS NIH HHS/United States ; DP1 MH100706/MH/NIMH NIH HHS/United States ; R01 DK097768/DK/NIDDK NIH HHS/United States ; 5R01DK097768-03/DK/NIDDK NIH HHS/United States ; GM10407/GM/NIGMS NIH HHS/United States
• Volume:
  60
• Issue:
  3
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:3ff567ecb2d9a4cb992a64851a991edcaf46b72c71c1aabf98263f60ba02b5d4
• Download URL:
  https://stacks.cdc.gov/view/cdc/37888/cdc_37888_DS1.pdf
• File Type:
  [PDF - 1.29 MB ]