Extended Data Fig. 8: Assessing trans cleavage of other RNA substrates by Ba1Cas12a3.
From: RNA-triggered Cas12a3 cleaves tRNA tails to execute bacterial immunity
(a) Comparing cleavage of different RNA substrates containing CCCCA. An activated Ba1Cas12a3 RNP was indicated with the indicated substrate, and fluorescence through the separation of the conjugated fluorophore (green circle) and quencher (gray circle) is measured over time for a range of substrate concentrations. Circles with nucleotides are colored to represent a mutation from C to U, a pyrimidine (red), or to G, a purine (yellow). The data from independent measurements (n = 3) were used to calculate catalytic efficiency (kcat/Km), with means ± standard error shown. (b) Comparing different truncated tRNA substrates with mutations to the cytosines in the 3′ CCA tail. Dotted curves and shaded regions represent the mean ± standard deviation of independently prepared and monitored reactions (n = 4).
