Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Record number 409333
Title Interference by clustered regularly interspaced short palindromic repeat (CRISPR) RNA is governed by a seed sequence
Author(s) Semenova, E.V.; Jore, M.M.; Westra, E.R.; Oost, J. van der; Brouns, S.J.J.
Source Proceedings of the National Academy of Sciences of the United States of America 108 (2011)25. - ISSN 0027-8424 - p. 10098 - 10103.
Department(s) Microbiological Laboratory
Publication type Refereed Article in a scientific journal
Publication year 2011
Keyword(s) immune-system - escherichia-coli - dna - defense - identification - prokaryotes - cleavage - bacteriophage - recognition - archaeon
Abstract Prokaryotic clustered regularly interspaced short palindromic repeat (CRISPR)/Cas (CRISPR-associated sequences) systems provide adaptive immunity against viruses when a spacer sequence of small CRISPR RNA (crRNA) matches a protospacer sequence in the viral genome. Viruses that escape CRISPR/Cas resistance carry point mutations in protospacers, though not all protospacer mutations lead to escape. Here, we show that in the case of Escherichia coli subtype CRISPR/Cas system, the requirements for crRNA matching are strict only for a seven-nucleotide seed region of a protospacer immediately following the essential protospacer-adjacent motif. Mutations in the seed region abolish CRISPR/Cas mediated immunity by reducing the binding affinity of the crRNA-guided Cascade complex to protospacer DNA. We propose that the crRNA seed sequence plays a role in the initial scanning of invader DNA for a match, before base pairing of the full-length spacer occurs, which may enhance the protospacer locating efficiency of the E. coli Cascade complex. In agreement with this proposal, single or multiple mutations within the protospacer but outside the seed region do not lead to escape. The relaxed specificity of the CRISPR/Cas system limits escape possibilities and allows a single crRNA to effectively target numerous related viruses
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