High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2013-09

AUTHORS

Yanfang Fu, Jennifer A Foden, Cyd Khayter, Morgan L Maeder, Deepak Reyon, J Keith Joung, Jeffry D Sander

ABSTRACT

Clustered, regularly interspaced, short palindromic repeat (CRISPR) RNA-guided nucleases (RGNs) have rapidly emerged as a facile and efficient platform for genome editing. Here, we use a human cell-based reporter assay to characterize off-target cleavage of CRISPR-associated (Cas)9-based RGNs. We find that single and double mismatches are tolerated to varying degrees depending on their position along the guide RNA (gRNA)-DNA interface. We also readily detected off-target alterations induced by four out of six RGNs targeted to endogenous loci in human cells by examination of partially mismatched sites. The off-target sites we identified harbored up to five mismatches and many were mutagenized with frequencies comparable to (or higher than) those observed at the intended on-target site. Our work demonstrates that RGNs can be highly active even with imperfectly matched RNA-DNA interfaces in human cells, a finding that might confound their use in research and therapeutic applications. More... »

PAGES

822-826

Journal

TITLE

Nature Biotechnology

ISSUE

9

VOLUME

31

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nbt.2623

    DOI

    http://dx.doi.org/10.1038/nbt.2623

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1009511624

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/23792628


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