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dc.contributor.authorHirano, Hisato
dc.contributor.authorHorii, Takuro
dc.contributor.authorKimura, Mika
dc.contributor.authorNakane, Takanori
dc.contributor.authorIshitani, Ryuichiro
dc.contributor.authorHatada, Izuho
dc.contributor.authorNishimasu, Hiroshi
dc.contributor.authorNureki, Osamu
dc.contributor.authorGootenberg, Jonathan S
dc.contributor.authorAbudayyeh, Omar Osama
dc.contributor.authorHsu, Patrick
dc.contributor.authorZhang, Feng
dc.date.accessioned2017-12-12T17:02:52Z
dc.date.available2017-12-12T17:02:52Z
dc.date.issued2016-02
dc.date.submitted2016-01
dc.identifier.issn0092-8674
dc.identifier.issn1097-4172
dc.identifier.urihttp://hdl.handle.net/1721.1/112719
dc.description.abstractSummary The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets complementary to the guide RNA and has been applied to programmable genome editing. Cas9-mediated cleavage requires a protospacer adjacent motif (PAM) juxtaposed with the DNA target sequence, thus constricting the range of targetable sites. Here, we report the 1.7 Å resolution crystal structures of Cas9 from Francisella novicida (FnCas9), one of the largest Cas9 orthologs, in complex with a guide RNA and its PAM-containing DNA targets. A structural comparison of FnCas9 with other Cas9 orthologs revealed striking conserved and divergent features among distantly related CRISPR-Cas9 systems. We found that FnCas9 recognizes the 5′-NGG-3′ PAM, and used the structural information to create a variant that can recognize the more relaxed 5′-YG-3′ PAM. Furthermore, we demonstrated that the FnCas9-ribonucleoprotein complex can be microinjected into mouse zygotes to edit endogenous sites with the 5′-YG-3′ PAM, thus expanding the target space of the CRISPR-Cas9 toolbox.en_US
dc.publisherElsevieren_US
dc.relation.isversionofhttp://dx.doi.org/10.1016/j.cell.2016.01.039en_US
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivs Licenseen_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/en_US
dc.sourcePMCen_US
dc.titleStructure and Engineering of Francisella novicida Cas9en_US
dc.typeArticleen_US
dc.identifier.citationHirano, Hisato et al. “Structure and Engineering of Francisella Novicida Cas9.” Cell 164, 5 (February 2016): 950–961 © 2016 Elsevier Incen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Brain and Cognitive Sciencesen_US
dc.contributor.departmentMcGovern Institute for Brain Research at MITen_US
dc.contributor.mitauthorGootenberg, Jonathan S
dc.contributor.mitauthorAbudayyeh, Omar Osama
dc.contributor.mitauthorHsu, Patrick
dc.contributor.mitauthorZhang, Feng
dc.relation.journalCellen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dc.date.updated2017-12-12T16:44:44Z
dspace.orderedauthorsHirano, Hisato; Gootenberg, Jonathan S.; Horii, Takuro; Abudayyeh, Omar O.; Kimura, Mika; Hsu, Patrick D.; Nakane, Takanori; Ishitani, Ryuichiro; Hatada, Izuho; Zhang, Feng; Nishimasu, Hiroshi; Nureki, Osamuen_US
dspace.embargo.termsNen_US
dc.identifier.orcidhttps://orcid.org/0000-0002-7979-3220
dc.identifier.orcidhttps://orcid.org/0000-0003-2782-2509
mit.licensePUBLISHER_CCen_US


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