Dynamics of CRISPR-Cas9 genome interrogation in living cells

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Science  13 Nov 2015:
Vol. 350, Issue 6262, pp. 823-826
DOI: 10.1126/science.aac6572

Genome editing with a Cas9 scalpel

The Cas9 nuclease forms the heart of the CRISPR-Cas genome editing system. Cas9 binds small guide RNAs that direct it to its target sites, where the nuclease either cleaves or binds to genomic DNA. Knight et al. used single-molecule imaging to track Cas9 in living cells. Cas9 searches the genome for its target sites using rapid threedimensional diffusion. It spends very little time binding to off-target sites, which explains the high accuracy of the CRISPRCas9 editing machine.

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