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Abstract

Prokaryotes acquire virus resistance by integrating short fragments of viral nucleic acid into clusters of regularly interspaced short palindromic repeats (CRISPRs). Here we show how virus-derived sequences contained in CRISPRs are used by CRISPR-associated (Cas) proteins from the host to mediate an antiviral response that counteracts infection. After transcription of the CRISPR, a complex of Cas proteins termed Cascade cleaves a CRISPR RNA precursor in each repeat and retains the cleavage products containing the virus-derived sequence. Assisted by the helicase Cas3, these mature CRISPR RNAs then serve as small guide RNAs that enable Cascade to interfere with virus proliferation. Our results demonstrate that the formation of mature guide RNAs by the CRISPR RNA endonuclease subunit of Cascade is a mechanistic requirement for antiviral defense.

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References and Notes

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We thank T. Verweij, C. G. J. van Houte, and M. R. Beijer for experimental contributions and T. Goosen (Hogeschool van Arnhem en Nijmegen BioCentre), M. J. Young (Montana State University), T. Bisseling, and W. M. de Vos(Wageningen University) for helpful discussions. We are grateful for receiving strains from the KEIO collection distributed by National BioResource Project (National Institute of Genetics, Japan). We thank U. Dobrindt (University of Würzburg) for sending genomic material of E. coli UTI89. This work was financially supported by a Vici grant from the Dutch Organization for Scientific Research (Nederlandse Organisatie voor Wetenschappelijk Onderzoek) and a Marie Curie grant from the European Union. M.L. was supported by the Wenner-Gren Foundations.

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Published In

Science
Volume 321 | Issue 5891
15 August 2008

Submission history

Received: 28 April 2008
Accepted: 1 July 2008
Published in print: 15 August 2008

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Notes

Supporting Online Material
www.sciencemag.org/cgi/content/full/321/5891/960/DC1
Materials and Methods
Figs. S1 to S4
Tables S1 to S3
References

Authors

Affiliations

Stan J. J. Brouns*
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, Netherlands.
Matthijs M. Jore*
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, Netherlands.
Magnus Lundgren
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, Netherlands.
Edze R. Westra
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, Netherlands.
Rik J. H. Slijkhuis
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, Netherlands.
Ambrosius P. L. Snijders
Biological and Environmental Systems, Department of Chemical and Process Engineering, University of Sheffield, Mappin Street, Sheffield S1 3JD, UK.
Mark J. Dickman
Biological and Environmental Systems, Department of Chemical and Process Engineering, University of Sheffield, Mappin Street, Sheffield S1 3JD, UK.
Kira S. Makarova
National Center for Biotechnology Information, National Library of Medicine, NIH, Bethesda, MD 20894, USA.
Eugene V. Koonin
National Center for Biotechnology Information, National Library of Medicine, NIH, Bethesda, MD 20894, USA.
John van der Oost [email protected]
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, Netherlands.

Notes

To whom correspondence should be addressed. E-mail: [email protected]

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