Thursday, May 29, 2014
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Genome editing with
dimeric, RNA-guided FokI-Cas9
In vitro studies suggest dimerization-dependent nucleases
could improve the specificity of clustered, regularly interspaced short palindromic
repeats (CRISPR)-based genome editing for
research and therapeutic applications. FokI nuclease, which requires
dimerization for DNA cleavage, was fused to a catalytically inactive version
of Cas9. In cultured human cells, expression of the dimeric editing tool and
guide RNAs cleaved the enhanced GFP reporter target and 11 of 12 target genes
without affecting any of 5 previously identified off-target sites for
wild-type Cas9. Also in cultured cells, expression of a distinct FokI-Cas9
dimeric nuclease with guide RNAs allowed targeting of 14 genomic sites with
lower efficiency but higher specificity than wild-type Cas9 or monomeric Cas9
nickases. Next steps include modifications to other aspects of CRISPR-based
technology including delivery and toxicity.
Published online May 29, 2014
Patent and licensing status
unavailable for findings from first study
For findings from second study, patent application filed by Harvard University, which is in
negotiations with Editas Medicine for licensing a
variety of CRISPR-related IP
Tsai, S.Q. et al. Nat.
Biotechnol.; published online April 25, 2014;
Contact: J. Keith Joung, Massachusetts General Hospital, Boston, Mass.
Guilinger, J.P. et al.
Nat. Biotechnol.; published online April 25, 2014;
Contact: David R. Liu, Harvard University, Cambridge, Mass.
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