Real-time observation of DNA recognition and rejection by the RNA-guided endonuclease Cas9

@inproceedings{Singh2016RealtimeOO,
  title={Real-time observation of DNA recognition and rejection by the RNA-guided endonuclease Cas9},
  author={Digvijay Singh and Samuel H Sternberg and Jingyi Fei and Jennifer A. Doudna and Taekjip Ha},
  booktitle={Nature communications},
  year={2016}
}
Binding specificity of Cas9-guide RNA complexes to DNA is important for genome-engineering applications; however, how mismatches influence target recognition/rejection kinetics is not well understood. Here we used single-molecule FRET to probe real-time interactions between Cas9-RNA and DNA targets. The bimolecular association rate is only weakly dependent on sequence; however, the dissociation rate greatly increases from <0.006 s(-1) to >2 s(-1) upon introduction of mismatches proximal to… CONTINUE READING