Figure 2: Cas9–RNA binding to DNA with proximal or distal mismatches.
From: Real-time observation of DNA recognition and rejection by the RNA-guided endonuclease Cas9
(a) A series of fully duplexed DNA targets with a varying number of mismatches (black segments) relative to the guide RNA. An x–ymm target has a contiguous mismatch running from position x to y relative to PAM. (b,c) FRET histograms of Cas9–RNA binding to DNA constructs carrying PAM-distal (b) and PAM-proximal (c) mismatches. The number of molecules for each histogram ranged from 568 to 3,053. [Cas9–RNA]=20 nM. (d) The fraction of Cas9–RNA-bound DNA molecules for different DNA targets. All the data shown in the figure are from independent experiments and error bars represent s.d. for n=3 (n=2 for few sets).
