Thursday, September 12, 2013
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Enhancing specificity of
RNA-guided clustered, regularly interspaced short palindromic repeats
(CRISPR)-based genome editing with paired guide RNAs
Enhancing the specificity
of CRISPR-based genome editing systems could improve its use in therapeutic
applications. CRISPR systems use small guide RNAs that pair with a target DNA
sequence and the CRISPR-associated protein 9 (Cas9) to excise target DNA. To
improve specificity, a system was designed that uses a Cas9 mutant enzyme
plus a pair of guide RNAs to excise a targeted DNA sequence. In cell lines,
this CRISPR-based editing system decreased off-target DNA excision by 50- to
1,000-fold compared with systems that use single guide RNAs. Next steps
include whole-genome sequencing studies to better characterize the
specificity of the Cas9 mutant and further improving the efficiency of the
system (see Crisper results for CRISPR, page 9).
SciBX 6(35); doi:10.1038/scibx.2013.978
Published online Sept. 12, 2013
Patent application filed;
available for licensing from the Broad
Institute of MIT and Harvard Office of Strategic Alliances
Ran, F.A. et al. Cell;
Aug. 29, 2013;
Contact: Feng Zhang, Broad Institute of MIT and Harvard, Cambridge,
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