Fig. 4

Comparison of gene silencing kinetics by crRNA and siRNA in a multiplexed setting. a U2OS-EGFP¹²-Cas9 cells were reverse transfected with a non-targeting control crRNA (NT), a combination of crRNAs targeting EGFP, PTEN, and KRAS (Combo), a non-targeting control siRNA pool (NT), or a combination of siRNAs targeting EGFP, PTEN, and KRAS (Combo). Protein expression was assessed by western blot at the indicated time points. b Quantification of protein levels from western blots in a. The decay of protein levels was fitted to a delayed one-phase decay model. c Parallel CRISPR transfection samples were collected at the indicated time points, and target site mutation was determined using TIDE. The decay of the fraction of WT allele was fitted to a delayed one-phase decay model. d Estimated half-lives of the EGFP, PTEN and KRAS WT allele and that of their proteins following CRISPR ablation and siRNA knockdown. Half-life was calculated using the fitted model in b and c