期刊
MOLECULAR CELL
卷 79, 期 3, 页码 416-+出版社
CELL PRESS
DOI: 10.1016/j.molcel.2020.06.022
关键词
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资金
- Centers for Excellence in Genomic Science of the National Institutes of Health (NIH) [RM1HG009490]
- National Science Foundation [1817593]
- Allen Distinguished Investigator Program through the Paul G. Allen Frontiers Group
- Somatic Cell Genome Editing Program of the Common Fund of the NIH [U01AI142817-02]
- NIH [RAI092531A]
- Innovative Genomics Institute
- Edmond J. Safra Center for Bioinformatics at Tel Aviv University
- Direct For Biological Sciences [1817593] Funding Source: National Science Foundation
- Div Of Molecular and Cellular Bioscience [1817593] Funding Source: National Science Foundation
CRISPR-Cas12c/d proteins share limited homology with Cas12a and Cas9 bacterial CRISPR RNA (crRNA)-guided nucleases used widely for genome editing and DNA detection. However, Cas12c (C2c3)- and Cas12d (CasY)-catalyzed DNA cleavage and genome editing activities have not been directly observed. We show here that a short-complementarity untranslated RNA (scoutRNA), together with crRNA, is required for Cas12d-catalyzed DNA cutting. The scoutRNA differs in secondary structure from previously described tracrRNAs used by CRISPR-Cas9 and some Cas12 enzymes, and in Cas12d-containing systems, scoutRNA includes a conserved five-nucleotide sequence that is essential for activity. In addition to supporting crRNA-directed DNA recognition, biochemical and cell-based experiments establish scoutRNA as an essential cofactor for Cas12c-catalyzed pre-crRNA maturation. These results define scoutRNA as a third type of transcript encoded by a subset of CRISPR-Cas genomic loci and explain how Cas12c/d systems avoid requirements for host factors including ribonuclease III for bacterial RNA-mediated adaptive immunity.
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