DNA targeting specificity of RNA-guided Cas9 nucleases.
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ABSTRACT:
SUBMITTER: Hsu PD
PROVIDER: S-EPMC3969858 | biostudies-literature | 2013 Sep
REPOSITORIES: biostudies-literature
Publications
DNA targeting specificity of RNA-guided Cas9 nucleases.
Hsu Patrick D PD Scott David A DA Weinstein Joshua A JA Ran F Ann FA Konermann Silvana S Agarwala Vineeta V Li Yinqing Y Fine Eli J EJ Wu Xuebing X Shalem Ophir O Cradick Thomas J TJ Marraffini Luciano A LA Bao Gang G Zhang Feng F
Nature biotechnology 20130721 9
The Streptococcus pyogenes Cas9 (SpCas9) nuclease can be efficiently targeted to genomic loci by means of single-guide RNAs (sgRNAs) to enable genome editing. Here, we characterize SpCas9 targeting specificity in human cells to inform the selection of target sites and avoid off-target effects. Our study evaluates >700 guide RNA variants and SpCas9-induced indel mutation levels at >100 predicted genomic off-target loci in 293T and 293FT cells. We find that SpCas9 tolerates mismatches between guid ...[more]
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