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. Author manuscript; available in PMC: 2019 Apr 5.
Published in final edited form as: Nat Struct Mol Biol. 2018 Apr 5;25(4):347–354. doi: 10.1038/s41594-018-0051-7

Figure 3. smFRET assay to investigate initial DNA unwinding upon binding of DNA to Cas9-RNA.

Figure 3.

(a) Schematic of smFRET assay for DNA unwinding by surface-tethered Cas9-RNA. (b) Representative single molecule fluorescence intensity time traces of donor (green) and acceptor (red) show abrupt signal increase upon DNA binding followed by FRET changes. (c) Time taken to go from initial binding to its first unwound state configuration (τunwinding) as denoted by a dark yellow line (a). Error bars represent standard deviation (s.d.) from n=2. n=1 in absence of error bar.