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dc.contributor.authorKleinstiver, Benjamin P.en_US
dc.contributor.authorPattanayak, Vikramen_US
dc.contributor.authorPrew, Michelle S.en_US
dc.contributor.authorTsai, Shengdar Q.en_US
dc.contributor.authorNguyen, Nhuen_US
dc.contributor.authorZheng, Zonglien_US
dc.contributor.authorJoung, J. Keithen_US
dc.date.accessioned2016-08-09T14:53:31Z
dc.date.issued2015en_US
dc.identifier.citationKleinstiver, Benjamin P., Vikram Pattanayak, Michelle S. Prew, Shengdar Q. Tsai, Nhu Nguyen, Zongli Zheng, and J. Keith Joung. 2015. “High-fidelity CRISPR-Cas9 variants with undetectable genome-wide off-targets.” Nature 529 (7587): 490-495. doi:10.1038/nature16526. http://dx.doi.org/10.1038/nature16526.en
dc.identifier.issn0028-0836en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:27822297
dc.description.abstractCRISPR-Cas9 nucleases are widely used for genome editing but can induce unwanted off-target mutations. Existing strategies for reducing genome-wide off-targets of the broadly used Streptococcus pyogenes Cas9 (SpCas9) are imperfect, possessing only partial or unproven efficacies and other limitations that constrain their use. Here we describe SpCas9-HF1, a high-fidelity variant harboring alterations designed to reduce non-specific DNA contacts. SpCas9-HF1 retains on-target activities comparable to wild-type SpCas9 with >85% of single-guide RNAs (sgRNAs) tested in human cells. Strikingly, with sgRNAs targeted to standard non-repetitive sequences, SpCas9-HF1 rendered all or nearly all off-target events undetectable by genome-wide break capture and targeted sequencing methods. Even for atypical, repetitive target sites, the vast majority of off-targets induced by SpCas9-HF1 were not detected. With its exceptional precision, SpCas9-HF1 provides an alternative to wild-type SpCas9 for research and therapeutic applications. More broadly, our results suggest a general strategy for optimizing genome-wide specificities of other RNA-guided nucleases.en
dc.language.isoen_USen
dc.relation.isversionofdoi:10.1038/nature16526en
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC4851738/pdf/en
dash.licenseLAAen_US
dc.titleHigh-fidelity CRISPR-Cas9 variants with undetectable genome-wide off-targetsen
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden
dc.relation.journalNatureen
dash.depositing.authorKleinstiver, Benjamin P.en_US
dc.date.available2016-08-09T14:53:31Z
dc.identifier.doi10.1038/nature16526*
dash.contributor.affiliatedPrew, Michelle
dash.contributor.affiliatedNguyen, Nhu
dash.contributor.affiliatedPattanayak, Vikram
dash.contributor.affiliatedKleinstiver, Benjamin
dash.contributor.affiliatedTsai, Shengdar Q.


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