(594g) Inducible Directed Evolution of Multigene Pathways in E. coli

Directed evolution is a powerful method for engineering biology in the absence of detailed sequence-function relationships. To enable directed evolution of complex phenotypes encoded by multigene pathways, we require large library sizes for DNA sequences >5-10kb in length, elimination of genomic hitchhiker mutations, and decoupling of diversification and screening steps. To meet these challenges, we developed Inducible Directed Evolution (IDE), which uses a temperate bacteriophage to package large plasmids and transfer them to naive cells after intracellular mutagenesis. Here, I will discuss the development of IDE, as well as its ability to quickly achieve improvements to several multigenic phenotypes, including carbon catabolism, pH tolerance, and biochemical production.