Mutation accumulation measurement for OrthoRep engineering and application toward TrpB evolution
We used directed evolution to engineer TP-DNAP1 from the OrthoRep in vivo mutagenesis system in yeast. High throughput sequencing was used to measure mutation rates to screen individual polymerase variants for elevated mutation rates and altered mutational preferences. We also used high throughput sequencing to examine the mutational landscape explored by TrpB evolution, and to measure TrpB fitness in high throughput.