Figure 4. Comparison of the linearization and nicking activities of SpyCas9^WT and SpyCas9^2Pro.

A) Analysis of cleavage pattern of SpyCas9^WT (left) and SpyCas9^2Pro (right) with a fully matched DNA substrate at 5 mM Mg²⁺ ions. B) Analysis of cleavage pattern of a mismatched (MM5) DNA substrate at 5 mM Mg²⁺ ions using SpyCas9^WT (left) and SpyCas9^2Pro (right). The average values for nicked (%), linear (%), and nicked + linear (%) (see Materials and Methods) are plotted against protein concentration. Data were obtained from three replications with a reaction time of 15 minutes and error bars represent SEM.