Using the hyperthermophile Pyrococcus furiosus, we have delineated several key steps in CRISPR (clustered regularly interspaced short palindromic repeats)–Cas (CRISPR-associated) invader defence pathways. P. furiosus has seven transcriptionally active CRISPR loci that together encode a total of 200 crRNAs (CRISPR RNAs). The 27 Cas proteins in this organism represent three distinct pathways and are primarily encoded in two large gene clusters. The Cas6 protein dices CRISPR locus transcripts to generate individual invader-targeting crRNAs. The mature crRNAs include a signature sequence element (the 5′ tag) derived from the CRISPR locus repeat sequence that is important for function. crRNAs are tailored into distinct species and integrated into three distinct crRNA–Cas protein complexes that are all candidate effector complexes. The complex formed by the Cmr [Cas module RAMP (repeat-associated mysterious proteins)] (subtype III-B) proteins cleaves complementary target RNAs and can be programmed to cleave novel target RNAs in a prokaryotic RNAi-like manner. Evidence suggests that the other two CRISPR–Cas systems in P. furiosus, Csa (Cas subtype Apern) (subtype I-A) and Cst (Cas subtype Tneap) (subtype I-B), target invaders at the DNA level. Studies of the CRISPR–Cas systems from P. furiosus are yielding fundamental knowledge of mechanisms of crRNA biogenesis and silencing for three of the diverse CRISPR–Cas pathways, and reveal that organisms such as P. furiosus possess an arsenal of multiple RNA-guided mechanisms to resist diverse invaders. Our knowledge of the fascinating CRISPR–Cas pathways is leading in turn to our ability to co-opt these systems for exciting new biomedical and biotechnological applications.
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December 2013
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Conference Article|
November 20 2013
The RNA- and DNA-targeting CRISPR–Cas immune systems of Pyrococcus furiosus
Rebecca M. Terns;
Rebecca M. Terns
1
*Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, U.S.A.
1Correspondence may be addressed to either author (emailrterns@bmb.uga.edu or mterns@bmb.uga.edu).
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Michael P. Terns
Michael P. Terns
1
*Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, U.S.A.
†Department of Genetics, University of Georgia, Athens, GA 30602, U.S.A.
‡Department of Microbiology, University of Georgia, Athens, GA 30602, U.S.A.
1Correspondence may be addressed to either author (emailrterns@bmb.uga.edu or mterns@bmb.uga.edu).
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Publisher: Portland Press Ltd
Received:
May 31 2013
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© The Authors Journal compilation © 2013 Biochemical Society
2013
Biochem Soc Trans (2013) 41 (6): 1416–1421.
Article history
Received:
May 31 2013
Citation
Rebecca M. Terns, Michael P. Terns; The RNA- and DNA-targeting CRISPR–Cas immune systems of Pyrococcus furiosus. Biochem Soc Trans 1 December 2013; 41 (6): 1416–1421. doi: https://doi.org/10.1042/BST20130056
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