This week in techniques



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Assays & screens

Quantification of clustered, regularly interspaced short palindromic repeats (CRISPR)-based editing system specificity

Quantification of off-target mutagenesis in CRISPR-based genome editing systems could help evaluate CRISPR systems for potential therapeutic applications and guide the development of CRISPR systems with increased accuracy. CRISPR systems use a small guide RNA, which pairs with a target DNA sequence and the CRISPR-associated protein (Cas9) to excise target DNA. The system recently has been used to rapidly engineer mice and cell lines carrying multiple mutations. A human cell-based GFP assay was used to quantify rates of off-target mutagenesis and found a range of 5.6%-125% off-target activity. Next steps include whole-genome mapping of off-target sites.

SciBX 6(29); doi:10.1038/scibx.2013.768
Published online Aug. 1, 2013

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Fu, Y. et al. Nat. Biotechnol.; published online June 23, 2013;
Contact: Jeffry D. Sander, Massachusetts General Hospital, Charlestown, Mass.

Contact: J. Keith Joung, same affiliation as above