Supplementary Figure 2: E histograms and Transition density plots for different DNA targets obtained from smFRET DNA-interrogation experiments at 20 nM Cas9–RNA. | Nature Structural & Molecular Biology

Supplementary Figure 2: E histograms and Transition density plots for different DNA targets obtained from smFRET DNA-interrogation experiments at 20 nM Cas9–RNA.

From: Mechanisms of improved specificity of engineered Cas9s revealed by single-molecule FRET analysis

Supplementary Figure 2

E histograms for (a) deCas9. (b) dCas9-HF1. (c) WT Cas9. (d) Transition density plots pictorialize the relative number of transitions between different FRET states, as identified by hidden Markov modeling. A small fraction of binding events showed rapid fluctuations between high and mid FRET states. The source of these fluctuations, also observed for WT Cas9(Singh et al., Nat Commun. 7, 12778, 2016), is unknown and dwells in these states were not included in the lifetime calculation. The number of PAM-distal mismatches (nPD) and PAM-proximal mismatches (nPP) are shown in cyan and orange, respectively.

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