Extended Data Fig. 6: Kinetic analysis of RBD#3L, RBD#4L and WT LwaCas13a.
From: Engineered LwaCas13a with enhanced collateral activity for nucleic acid detection
a–c, Progress curves of cleaved reporters versus time. Data were plotted as mean ± s.e.m. from two independent experiments each with four technical replicates. The RNP complex of RBD#3L, RBD#4L, and WT was incubated with 50 pM of the targets (T1) for 10 min at 37 °C prior to cleavage reaction using 6400, 3200, 1600, 800, 400, 200, 100 and 50 nM of the U11 substrates. The initial velocities (within 600 s) of each reaction were obtained by linear regression. d, Initial velocities from two independent experiments each with four technical replicates obtained from a–c were plotted as mean ± s.e.m. versus the U11 reporter concentration and fitted to a Michaelis-Menten curve. Summary of kinetic parameters are listed in Supplementary Table 4.
