Fig. 4.

Extrapolation of the dCas9 dynamic model to assess single target cleavage by Cas9. a The proposed model surrounding dCas9 interaction with the obtained kinetic rates. Free dCas9 (yellow) in the cytoplasm interact with PAM sequences (5’-NGG-3’) on average every 25 ms. If the PAM is not in front of a target sequence (red), only PAM-screening will occur for on average 17 ms. If the PAM happens to be in front of a target, the dCas9 will be target-bound (purple). We extend this model to predict Cas9 cleavage under conditions where target-bound Cas9 will always cleave the target DNA. b Calculated predicted probability that a single target in the L. lactis genome is cleaved after a certain period of time with a certain cellular Cas9 copy number, based on the model shown in a. Error bars indicate standard deviation calculated from iterations of the model