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New techniques improve specificity of CRISPR/CAS9 genome editing tools

To overcome the off-target mutations that commonly occur with CRISPR/Cas9 genome editing methods, researchers at and Massachusetts General Hospital have developed two strategies that greatly improve the specificity of RNA-guided nucleases for the DNA region targeted to be cut and repaired. A description of these new techniques and their successful use to modify human cancer cells and embryonic stem cells is described in a special issue on genome editing in , a peer-reviewed journal from , , publishers.

In the special issue led by Guest Editor Feng Zhang, PhD, Broad Institute of MIT and Harvard, and , MIT (Cambridge, MA), ?Nicolas Wyvekens, Ved Topkar, Cyd Khayter, J. Keith Joung, and Shengdar Tsai present their work in the article “Dimeric CRISPR RNA-Guided FokI-dCas9 Nucleases Directed by Truncated gRNAs for Highly Specific Genome Editing.” They introduce two new strategies to reduce the off-target effects of current methods: the use of truncated guide RNA molecules (gRNAs), creating shorter binding sites between the gRNAs and targeted DNA regions; and the addition of a FokI domain to the Cas9 protein, resulting in the formation of a nuclease dimer instead of monomer (the RNA-guided FokI-dCas9 Nuclease, or RFN).

Research reported in this publication was supported by the National Institutes of Health under Award Numbers DP1 GM105378, P50 HG005550, R01 AR063070, and F32 GM105189

Research: Dimeric CRISPR RNA-Guided FokI-dCas9 Nucleases Directed by Truncated gRNAs for Highly Specific Genome Editing, Wyvekens Nicolas, Topkar Ved V., Khayter Cyd, Joung J. Keith, and Tsai Shengdar Q, Human Gene Therapy, doi:10.1089/hum.2015.084, published online 12 June 2015.

Source

Source: Mary Ann Liebert, Inc., Publishers