Extended Data Figure 3. Kinetic analysis of transitions between active and inactive states of the HNH domain.
a, Representative time traces (top), transition density plots (TDPs, middle) and rates of the transitions in TDPs (bottom) for SpCas9-HF1 with on-target DNA (left), eSpCas9(1.1) with on-target DNA (middle) and eSpCas9(1.1) with 20-20 bp mm DNA (right); mean and s.e.m. shown; n = 107, 24, and 74 individual molecules, respectively. The percentage of molecules that show at least one such transitions was 36%, 7% and 29% for SpCas9-HF1 with on-target, eSpCas9(1.1) with on-target and eSpCas9(1.1) with 20-20 bp mm DNA, respectively. Kinetics analysis of other cases (SpCas9-HF1 and eSpCas9(1.1) bound to other off-target substrates, and HypaCas9 bound to on- and off-target substrates) is not shown, because the percentage of molecules that show at least one such transitions was less than 3%. b, Comparison of on-target transition rates for WT SpCas9, SpCas9-HF1, and eSpCas9(1.1); mean and s.e.m. shown; n = 51, 107 and 24 individual molecules, respectively. Transition rates for WT SpCas9 collected from ref. 10.