Selected applications of base editor-based targeted random mutagenesis (BE-TRM) tools for directed evolution (DE) of specific loci in bacteria, yeast, plants, and human cells. In these studies, the DE strategy was employed to evolve the rpoB in Escherichia coli, PfDHFR in yeasts, OsACC in rice plants, and both hTDR and promoter of HBG1/2 gene in human cells. To induce hypermutations in the targeted DNA regions, appropriate BE-TRM tools were utilized. The resulting mutants were then screened to identify gain-of-function variants. Further details regarding the genes, gain-of-function effects, and tool design can be found in the main text and Table 1.
