Scalable, Continuous Evolution of Genes at Mutation Rates above Genomic Error Thresholds.

ABSTRACT: Directed evolution is a powerful approach for engineering biomolecules and understanding adaptation. However, experimental strategies for directed evolution are notoriously labor intensive and low throughput, limiting access to demanding functions, multiple functions in parallel, and the study of molecular evolution in replicate. We report OrthoRep, an orthogonal DNA polymerase-plasmid pair in yeast that stably mutates ∼100,000-fold faster than the host genome in vivo, exceeding the error threshold of genomic replication that causes single-generation extinction. User-defined genes in OrthoRep continuously and rapidly evolve through serial passaging, a highly straightforward and scalable process. Using OrthoRep, we evolved drug-resistant malarial dihydrofolate reductases (DHFRs) in 90 indep

SUBMITTER: Ravikumar A 

PROVIDER: S-EPMC6343851 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature