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A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).
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2012 (English)In: Science, ISSN 0036-8075, E-ISSN 1095-9203, Vol. 337, no 6096, 816-821 p.Article in journal (Refereed) Published
Abstract [en]

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. We show here that in a subset of these systems, the mature crRNA that is base-paired to trans-activating crRNA (tracrRNA) forms a two-RNA structure that directs the CRISPR-associated protein Cas9 to introduce double-stranded (ds) breaks in target DNA. At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand. The dual-tracrRNA:crRNA, when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage. Our study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.

Place, publisher, year, edition, pages
2012. Vol. 337, no 6096, 816-821 p.
National Category
Microbiology in the medical area
Identifiers
URN: urn:nbn:se:umu:diva-59521DOI: 10.1126/science.1225829ISI: 000307535600036OAI: oai:DiVA.org:umu-59521DiVA: diva2:552965
Available from: 2012-09-17 Created: 2012-09-17 Last updated: 2018-01-12Bibliographically approved

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Chylinski, KrzysztofFonfara, InesCharpentier, Emmanuelle
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Molecular Infection Medicine Sweden (MIMS)Umeå Centre for Microbial Research (UCMR)Department of Molecular Biology (Faculty of Medicine)
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