RNA-Guided RNA Cleavage by a CRISPR RNA-Cas Protein Complex

By Caryn R. Hale, Peng Zhao, S. Olson, M. Duff, Brenton R. Graveley, L. Wells, R. Terns, M. Terns. Cell, 2009

1,043 citations recorded by the source corpus.

SUMMARY Compelling evidence indicates that the CRISPR-Cas system protects prokaryotes from viruses and other potential genome invaders. This adaptive prokaryotic immune system arises from the clustered regularly interspaced short palindromic repeats (CRISPRs) found in prokaryotic genomes, which harbor short invader-derived sequences, and the CRISPR-associated (Cas) protein-coding genes. Here we have identified a CRISPR-Cas effector complex that is comprised of small invader-targeting RNAs from the CRISPR loci (termed prokaryotic silencing (psi)RNAs) and the RAMP module (or Cmr) Cas proteins. The psiRNA-Cmr protein complexes cleave complementary target RNAs at a fixed distance from the 3' end of the integral psiRNAs. In Pyrococcus furiosus, psiRNAs occur in two size forms that share a common 5' sequence tag but have distinct 3' ends that direct cleavage of a given target RNA at two distinct sites. Our results indicate that prokaryotes possess a unique RNA silencing system that functions by homology-dependent cleavage of invader RNAs.

Subfields: Ecology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/507d7ed038a2c1677a3aa02caeb483093cefb9a8

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Citations in the corpus

Biology and Applications of CRISPR Systems: Harnessing Nature's Toolbox for Genome Engineering. (2016, neutral): “Early in vivo genetic experiments suggested Csm targeted DNA, while in vitro studies of Cmr showed binding and cleavage activity against RNA only (Hale et al., 2009; Marraffini and Sontheimer, 2008), leading to a model wherein the two subtypes had evolved distinct and complementary substrate prefere”
The new frontier of genome engineering with CRISPR-Cas9 (2014, neutral): “The type I and type III systems use a large complex of Cas proteins for crRNA-guided targeting ( 47 , 59 – 63 ).”
CRISPR interference: RNA-directed adaptive immunity in bacteria and archaea (2010, background): “coli , cleavage occurs at a specific site 8 nt upstream of the spacer, and the resulting crRNAs are then trimmed in vivo at their 3′ but not 5′ end”
Identifying and visualizing functional PAM diversity across CRISPR-Cas systems (2016, background): “…their 20–30 nucleotide spacer region to base pair with complementary protospacer sequences, leading the system’s Cas effector proteins to cleave or degrade these targets (Garneau et al., 2010; Gasiunas et al., 2012; Hale et al., 2009; Jinek et al., 2013; Jore et al., 2011; Westra et al., 2012).”
CRISPR-Cas systems: prokaryotes upgrade to adaptive immunity (2014, background): “(C) In type III systems, Cas6 cleaves the pre-crRNA to generate int-crRNAs that are incorporated into a Cmr/Cas10 or Csm/Cas10 complex, where further maturation occurs through the trimming of 30 end sequences (Hale et al., 2009, 2012).”
Development and Applications of CRISPR-Cas9 for Genome Engineering (2014, neutral): “Cell 157, June 5, 2014 ª2014 Elsevier Inc. 1265 2008), although later investigation of a different type III-B system from Pyrococcus furiosus also revealed crRNA-directed RNA cleavage activity (Hale et al., 2009, 2012).”
Transcriptome engineering with RNA-targeting Type VI-D CRISPR effectors (2018, neutral): “Although some class 1 CRISPR systems can target RNA (Hale et al., 2009; Jiang et al., 2016; Kazlauskiene et al., 2017; Niewoehner et al., 2017; Samai et al., 2015), type VI systems have been recently described as the only known single-effector CRISPR nucleases that exclusively target RNA (Abudayyeh ”