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. 2019 Nov 13;5(11):eaaw9807. doi: 10.1126/sciadv.aaw9807

Fig. 1. Experimental configuration for single-molecule unzipping experiments and dCas9/sgRNA/DNA interaction mapping.

Fig. 1

(A) Cartoon illustrating the single-molecule DNA unzipping experiment used for the detection of the interactions between dCas9/sgRNA and DNA. The sequence of the steps of the unzipping of the DNA template is numbered as illustrated. (B and E) Representative traces of forward (black) and reverse unzipping (blue) in the presence of dCas9 showing the force versus number of base pairs unzipped. The naked DNA unzipping signatures are also presented for comparison (gray). The light-yellow rectangle shows the expected Cas9 binding position. Red arrows indicate the force peaks. Insets: Zoom in of the regions with increases in force. (C and F) Histogram of the disruption forces of dCas9/sgRNA. The data and their Gaussian fits are shown. (D and G) Histogram of the positions of dCas9/sgRNA and target DNA interactions along the DNA sequence. The data and their Gaussian fits are shown. The black and blue triangles indicate the pre- and post-PAM interactions, respectively.