Cas9 endonuclease and off-target activity

Abstract

Genome Sequencing The RNA-guided endonuclease Cas9 has brought about a revolution in genetic engineering and genome editing. However, issues with the system include the off-target activity of the endonuclease and promiscuous cleavage, making accuracy in vivo a challenge. Dagdas et al. used smFRET (single-molecule Forster resonance energy transfer) to gain a better molecular understanding of this process, which should enhance the specificity of DNA targeting by Cas9. Sci. Adv. 10.1126/sciadv.aao0027 (2017).