Fig. 4. CPP can be conjugated to Cas9 via DEC and releases active Cas9 under cellular concentrations of GSH. (a) Structure of CPP–DEC. (b) CPP–DEC can be conjugated to Cas9 and releases active Cas9 in the presence of cellular concentrations of GSH (10 mM). (c) Nuclease activity of the Cas9 RNP after conjugation with various mole ratios of CPP–DEC (X-axis). Red line represents nuclease activity after incubation with GSH (10 mM). (d) Kinetics of CPP release from Cas9 after addition of 10 mM GSH (0, 1, 5, 10, 20, 30 minutes), the fluorescent bands show the time dependent release of CPP from Cas9–CPP.
