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Nat Rev Genet. 2016 May;17(5):300-12. doi: 10.1038/nrg.2016.28.

Defining and improving the genome-wide specificities of CRISPR-Cas9 nucleases.

Author information

1
Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, 149 13th Street, Charlestown, Massachusetts 02129, USA; and the Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA.

Abstract

CRISPR-Cas9 RNA-guided nucleases are a transformative technology for biology, genetics and medicine owing to the simplicity with which they can be programmed to cleave specific DNA target sites in living cells and organisms. However, to translate these powerful molecular tools into safe, effective clinical applications, it is of crucial importance to carefully define and improve their genome-wide specificities. Here, we outline our state-of-the-art understanding of target DNA recognition and cleavage by CRISPR-Cas9 nucleases, methods to determine and improve their specificities, and key considerations for how to evaluate and reduce off-target effects for research and therapeutic applications.

PMID:
27087594
DOI:
10.1038/nrg.2016.28
[Indexed for MEDLINE]

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