Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions

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dc.contributor.ISNI0000 0001 2535 9739 (Bleris, L)en_US
dc.contributor.authorLi, Yien_US
dc.contributor.authorMendiratta, Saurabhen_US
dc.contributor.authorEhrhardt, Kristinaen_US
dc.contributor.authorKashyap, Nehaen_US
dc.contributor.authorWhite, Michael A.en_US
dc.contributor.authorBleris, Leonidasen_US
dc.date.accessioned2016-09-23T20:42:21Z
dc.date.available2016-09-23T20:42:21Z
dc.date.created2016-01-20en_US
dc.date.issued2016-01-20en_US
dc.description.abstractCRISPR/Cas9 is an enabling RNA-guided technology for genome targeting and engineering. An acute DNA binding constraint of the Cas9 protein is the Protospacer Adjacent Motif (PAM). Here we demonstrate that the PAM requirement can be exploited to specifically target single-nucleotide heterozygous mutations while exerting no aberrant effects on the wildtype alleles. Specifically, we target the heterozygous G13A activating mutation of KRAS in colorectal cancer cells and we show reversal of drug resistance to a MEK small-molecule inhibitor. Our study introduces a new paradigm in genome editing and therapeutic targeting via the use of gRNA to guide Cas9 to a desired protospacer adjacent motif.en_US
dc.description.sponsorship"This work was funded by the US National Institutes of Health (NIH) grants GM096271, CA170018, the US National Science Foundation (NSF) CAREER grant 1351354, the University of Texas at Dallas, and Robert Welch Foundation (I-1414).en_US
dc.identifier.bibliographicCitationLi, Yi, Saurabh Mendiratta, Kristina Ehrhardt, Neha Kashyap, et al. 2016. "Exploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventions." PLOS One 11(1), doi:10.1371/journal.pone.0144970.en_US
dc.identifier.issn1932-6203en_US
dc.identifier.issue1en_US
dc.identifier.urihttp://hdl.handle.net/10735.1/5054
dc.identifier.volume11en_US
dc.relation.urihttp://dx.doi.org/10.1371/journal.pone.0144970
dc.relation.urihttp://dx.doi.org/10.1371/journal.pone.0144970en_US
dc.rightsCC BY 4.0 (Attribution)en_US
dc.rights©2016 The Authorsen_US
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en_US
dc.source.journalPLOS Oneen_US
dc.subjectDNAen_US
dc.subjectKRAS protein, humanen_US
dc.subjectCas9 protein, Francisella novicidaen_US
dc.subjectCodonen_US
dc.subjectPlasmidsen_US
dc.subjectMutationen_US
dc.subjectRNA, Guideen_US
dc.subjectRNA, Messengeren_US
dc.titleExploiting the CRISPR/Cas9 PAM Constraint for Single-Nucleotide Resolution Interventionsen_US
dc.type.genrearticleen_US

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