作者
Ekaterina Semenova, Matthijs M Jore, Kirill A Datsenko, Anna Semenova, Edze R Westra, Barry Wanner, John Van Der Oost, Stan JJ Brouns, Konstantin Severinov
发表日期
2011/6/21
期刊
Proceedings of the National Academy of Sciences
卷号
108
期号
25
页码范围
10098-10103
出版商
National Academy of Sciences
简介
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 …
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E Semenova, MM Jore, KA Datsenko, A Semenova… - Proceedings of the National Academy of Sciences, 2011