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dc.contributor.authorDugar, Gaurav
dc.contributor.authorLeenay, Ryan T
dc.contributor.authorEisenbart, Sara K
dc.contributor.authorBischler, Thorsten
dc.contributor.authorAul, Belinda U
dc.contributor.authorBeisel, Chase L
dc.contributor.authorSharma, Cynthia M
dc.date.accessioned2019-10-16T12:56:46Z
dc.date.available2019-10-16T12:56:46Z
dc.date.issued2018-03-01
dc.identifier.citationMol Cell. 2018 Mar 1;69(5):893-905.e7. doi: 10.1016/j.molcel.2018.01.032.en_US
dc.identifier.issn1097-4164
dc.identifier.pmid29499139
dc.identifier.doi10.1016/j.molcel.2018.01.032
dc.identifier.urihttp://hdl.handle.net/10033/621980
dc.description.abstractCas9 nucleases naturally utilize CRISPR RNAs (crRNAs) to silence foreign double-stranded DNA. While recent work has shown that some Cas9 nucleases can also target RNA, RNA recognition has required nuclease modifications or accessory factors. Here, we show that the Campylobacter jejuni Cas9 (CjCas9) can bind and cleave complementary endogenous mRNAs in a crRNA-dependent manner. Approximately 100 transcripts co-immunoprecipitated with CjCas9 and generally can be subdivided through their base-pairing potential to the four crRNAs. A subset of these RNAs was cleaved around or within the predicted binding site. Mutational analyses revealed that RNA binding was crRNA and tracrRNA dependent and that target RNA cleavage required the CjCas9 HNH domain. We further observed that RNA cleavage was PAM independent, improved with greater complementarity between the crRNA and the RNA target, and was programmable in vitro. These findings suggest that C. jejuni Cas9 is a promiscuous nuclease that can coordinately target both DNA and RNA.en_US
dc.publisherElsevier/ Cel Pressen_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subjectCRISPRen_US
dc.subjectCampylobacter jejunien_US
dc.subjectCas9en_US
dc.subjectRIP-seqen_US
dc.subjectRNA binding proteinsen_US
dc.subjectRNA cleavageen_US
dc.subjectcrRNAen_US
dc.subjectgenome editingen_US
dc.subjectnon-coding RNAen_US
dc.subjectpost-transcriptional regulationen_US
dc.titleCRISPR RNA-Dependent Binding and Cleavage of Endogenous RNAs by the Campylobacter jejuni Cas9.en_US
dc.typeArticleen_US
dc.contributor.departmentHIRI, Helmholtz-Institut für RNA-basierte Infektionsforschung, Josef-Shneider Strasse 2, 97080 Würzburg, Germany.en_US
dc.identifier.journalMolecular Cellen_US
refterms.dateFOA2019-10-16T12:56:47Z
dc.source.journaltitleMolecular cell


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