VEGAS as a Platform for Facile Directed Evolution in Mammalian Cells.

ABSTRACT: Directed evolution, artificial selection toward designed objectives, is routinely used to develop new molecular tools and therapeutics. Successful directed molecular evolution campaigns repeatedly test diverse sequences with a designed selective pressure. Unicellular organisms and their viral pathogens are exceptional for this purpose and have been used for decades. However, many desirable targets of directed evolution perform poorly or unnaturally in unicellular backgrounds. Here, we present a system for facile directed evolution in mammalian cells. Using the RNA alphavirus Sindbis as a vector for heredity and diversity, we achieved 24-h selection cycles surpassing 10^-3 mutations per base. Selection is achieved through genetically actuated sequences internal to the host cell, t

SUBMITTER: English JG 

PROVIDER: S-EPMC6660416 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature