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dc.contributor.authorZebec, Z.
dc.contributor.authorManica, A.
dc.contributor.authorZhang, J.
dc.contributor.authorWhite, M.F.
dc.contributor.authorSchleper, C.
dc.date.accessioned2014-07-09T11:31:01Z
dc.date.available2014-07-09T11:31:01Z
dc.date.issued2014-04-01
dc.identifier.citationZebec , Z , Manica , A , Zhang , J , White , M F & Schleper , C 2014 , ' CRISPR-mediated targeted mRNA degradation in the archaeon Sulfolobus solfataricus ' Nucleic Acids Research , vol. 42 , no. 8 , pp. 5280-5288 . https://doi.org/10.1093/nar/gku161en
dc.identifier.issn0305-1048
dc.identifier.otherPURE: 122125202
dc.identifier.otherPURE UUID: d7a6d5d3-e0ef-49e7-98b9-95bead1e9eb7
dc.identifier.otherScopus: 84899048370
dc.identifier.otherWOS: 000336092300052
dc.identifier.otherORCID: /0000-0003-1543-9342/work/47136121
dc.identifier.urihttp://nar.oxfordjournals.org/content/42/8/5280en
dc.descriptionEuropean SulfoSYS-project [SysMo P–N-01-09-23] and grant [9P23000 and P25369] by the Austrian Research fund (to C.S.) and by grant [BB/K000314/1] from the Biotechnology and Biological Sciences Research Council (to M.F.W.). Funding for open access charge: Austrian Science Fund.en
dc.description.abstractThe recently discovered clustered regularly interspaced short palindromic repeat (CRISPR)-mediated virus defense represents an adaptive immune system in many bacteria and archaea. Small CRISPR RNAs cause cleavage of complementary invading nucleic acids in conjunction with an associated protein or a protein complex. Here, we show CRISPR-mediated cleavage of mRNA from an invading virus in the hyperthermophilic archaeon Sulfolobus solfataricus. More than 40% of the targeted mRNA could be cleaved, as demonstrated by quantitative polymerase chain reaction. Cleavage of the mRNA was visualized by northern analyses and cleavage sites were mapped. In vitro, the same substrates were cleaved by the purified CRISPR-associated CMR complex from Sulfolobus solfataricus. The in vivo system was also re-programmed to knock down mRNA of a selected chromosomal gene (β-galactosidase) using an artificial miniCRISPR locus. With a single complementary spacer, ∼50% reduction of the targeted mRNA and of corresponding intracellular protein activity was achieved. Our results demonstrate in vivo cleavage of mRNA in a prokaryote mediated by small RNAs (i.e. analogous to RNA interference in eukaryotes) and the re-programming of the system to silence specific genes of interest.
dc.format.extent9
dc.language.isoeng
dc.relation.ispartofNucleic Acids Researchen
dc.rights© The Author(s) 2014. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.en
dc.subjectQ Scienceen
dc.subject.lccQen
dc.titleCRISPR-mediated targeted mRNA degradation in the archaeon Sulfolobus solfataricusen
dc.typeJournal articleen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews.School of Biologyen
dc.contributor.institutionUniversity of St Andrews.Biomedical Sciences Research Complexen
dc.identifier.doihttps://doi.org/10.1093/nar/gku161
dc.description.statusPeer revieweden


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