RNA-dependent RNA targeting by CRISPR-Cas9
- University of California, Berkeley, United States
- Sandia National Laboratories, United States
- Cited 7
- Views 8,491
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Abstract
Double-stranded DNA (dsDNA) binding and cleavage by Cas9 is a hallmark of type II CRISPR-Cas bacterial adaptive immunity. All known Cas9 enzymes are thought to recognize DNA exclusively as a natural substrate, providing protection against DNA phage and plasmids. Here we show that Cas9 enzymes from both subtypes II-A and II-C can recognize and cleave single-stranded RNA (ssRNA) by an RNA-guided mechanism that is independent of a protospacer-adjacent motif (PAM) sequence in the target RNA. RNA-guided RNA cleavage is programmable and site-specific, and we find that this activity can be exploited to reduce infection by single-stranded RNA phage in vivo. We also demonstrate that Cas9 can direct PAM-independent repression of gene expression in bacteria. These results indicate that a subset of Cas9 enzymes have the ability to act on both DNA and RNA target sequences, and suggest the potential for use in programmable RNA targeting applications.
Article and author information
Author details
Jennifer A Doudna
Funding
Howard Hughes Medical Institute
- Rachel M Torrez
- Jennifer A Doudna
National Science Foundation (MCB-1244557)
- Steven C Strutt
- Jennifer A Doudna
Paul Allen Frontiers Science Program
- Jennifer A Doudna
Laboratory Directed Research and Development (U.S. Department of Energy's National Nuclear Security Administration under contract DE-NA0003525)
- Oscar A Negrete
German Academic Exchange Program
- Emine Kaya
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Rodolphe Barrangou, Reviewing Editor, North Carolina State University, United States
Publication history
- Received: October 12, 2017
- Accepted: January 3, 2018
- Accepted Manuscript published: January 5, 2018 (version 1)
- Version of Record published: February 2, 2018 (version 2)
Copyright
© 2018, Strutt et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Further reading
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- Biochemistry and Chemical Biology
- Structural Biology and Molecular Biophysics
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- Biochemistry and Chemical Biology
- Microbiology and Infectious Disease
