Actions for RNA-dependent RNA targeting by CRISPR-Cas9 [electronic resource].

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RNA-dependent RNA targeting by CRISPR-Cas9 [electronic resource].

Published
Washington, D.C. : United States. National Nuclear Security Administration, 2018.
Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy
Physical Description
Article numbers: e32,724 : digital, PDF file
Additional Creators
Lawrence Berkeley National Laboratory, United States. National Nuclear Security Administration, National Science Foundation (U.S.), United States. Department of Energy. Office of Fissile Materials Disposition, University of California, Berkeley, National Institutes of Health (U.S.), and United States. Department of Energy. Office of Scientific and Technical Information
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Summary
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. In conclusion, 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.
Report Numbers
E 1.99:1433112
Subject(s)
Note
Published through SciTech Connect.
01/05/2018.
"ark:/13030/qt1hz8w9tx"
eLife 7 none ISSN 2050-084X AM
Steven C. Strutt; Rachel M. Torrez; Emine Kaya; Oscar A. Negrete; Jennifer A. Doudna.
Funding Information
AC02-05CH11231
MCB-1244557
NA0003525
S10RR029668
S10RR027303
View MARC record | catkey: 23497775