Fig. 5. Illustration of protein structures and experimental results for multisite mutants, comparing their properties against wild-type proteins.
The protein structures and experimental results for multisite mutants of LbCas12a (predicted by Alphafold2) (A and C) and T7 RNA polymerase (PDB ID: 1MSW) (B and D). The functional domains of each protein are depicted beneath their structural diagrams. In (C) and (D), a red dashed line indicates the normalized activity level of the wild type. The activities of the five most thermostable mutants are marked with red dots to facilitate direct comparison with the wild type. The thermal stability (Tm) of the wild type is represented by a yellow dashed line for LbCas12a and a blue dashed line for T7 RNA polymerase. R1 and R2 represent the first and the second rounds of multipoint combination, respectively, with the numbers following them indicating the mutant indices.