Genome-Wide Assessment of Efficiency and Specificity in CRISPR/Cas9 Mediated Multiple Site Targeting in Arabidopsis

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dc.contributor.authorPeterson, Brenda A.en
dc.contributor.authorHaak, David C.en
dc.contributor.authorNishimura, Marc T.en
dc.contributor.authorTeixeira, Paulo J. P. L.en
dc.contributor.authorJames, Sean R.en
dc.contributor.authorDangl, Jeffery L.en
dc.contributor.authorNimchuk, Zachary L.en
dc.contributor.departmentBiological Sciencesen
dc.contributor.departmentSchool of Plant and Environmental Sciencesen
dc.date.accessioned2017-01-05T10:39:29Zen
dc.date.available2017-01-05T10:39:29Zen
dc.date.issued2016-09-13en
dc.description.abstractSimultaneous multiplex mutation of large gene families using Cas9 has the potential to revolutionize agriculture and plant sciences. The targeting of multiple genomic sites at once raises concerns about the efficiency and specificity in targeting. The model Arabidopsis thaliana is widely used in basic plant research. Previous work has suggested that the Cas9 off-target rate in Arabidopsis is undetectable. Here we use deep sequencing on pooled plants simultaneously targeting 14 distinct genomic loci to demonstrate that multiplex targeting in Arabidopsis is highly specific to on-target sites with no detectable off-target events. In addition, chromosomal translocations are extremely rare. The high specificity of Cas9 in Arabidopsis makes this a reliable method for clean mutant generation with no need to enhance specificity or adopt alternate Cas9 variants.en
dc.description.versionPublished versionen
dc.format.extent? - ? (11) page(s)en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1371/journal.pone.0162169en
dc.identifier.issn1932-6203en
dc.identifier.issue9en
dc.identifier.urihttp://hdl.handle.net/10919/73949en
dc.identifier.volume11en
dc.language.isoenen
dc.publisherPLOSen
dc.relation.urihttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000383681000015&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1en
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectchromosomal translocationsen
dc.subjecthigh-frequencyen
dc.subjectthalianaen
dc.subjectcas9en
dc.subjectmutagenesisen
dc.subjectnucleasesen
dc.subjectvisualizationen
dc.subjectendonucleaseen
dc.subjectpeptidesen
dc.subjecttalensen
dc.titleGenome-Wide Assessment of Efficiency and Specificity in CRISPR/Cas9 Mediated Multiple Site Targeting in Arabidopsisen
dc.title.serialPLOS ONEen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciencesen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/CALS T&R Facultyen
pubs.organisational-group/Virginia Tech/Agriculture & Life Sciences/Plant Pathology, Physiology, & Weed Scienceen
pubs.organisational-group/Virginia Tech/All T&R Facultyen

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