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Dynamics of CRISPR-Cas9 genome interrogation in living cells

2015; American Association for the Advancement of Science; Volume: 350; Issue: 6262 Linguagem: Inglês

10.1126/science.aac6572

1095-9203

Spencer C. Knight, Liangqi Xie, Wulan Deng, Benjamin Guglielmi, Lea B. Witkowsky, Lana Bosanac, Elisa Zhang, Mohamed El Beheiry, Jean‐Baptiste Masson, Maxime Dahan, Zhe Liu, Jennifer A. Doudna, Robert Tjian,

Genetics, Aging, and Longevity in Model Organisms

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. Science , this issue p. 823