Fig. 3.

sptPALM of dCas9-PAmCherry2 in pTarget L. lactis shows target-binding behaviour of dCas9. a Identified tracks in individual pTarget L. lactis cells. Tracks are colour-coded based on their diffusion coefficient. Three separate cells are shown with increasing dCas9 concentration. Blue dotted outline is an indication for the cell membrane. Scale bars represent 500 nm. b Diffusion coefficient histograms (light blue) are fitted (dark blue line) with a combination of the respective fit of pNonTarget L. lactis cells (green line), along with a single globally fitted population corresponding to target-bound dCas9 (purple) at 0.38 ± 0.04 µm²/s (mean ± standard deviation). c Left: The population size of the plasmid-bound dCas9 decreases as a function of the cellular dCas9 copy number. The error bar of the measurement is based on the 95% confidence interval determined by bootstrapping; the solid line is a model fit with 20 plasmids, with a 95% confidence interval determined by repeating the model simulation. Right: Occupancy of DNA targets by dCas9 based on 20 target plasmids (100 DNA target sites), based on the same data as presented in the left figure. Source data are provided as a Source Data file