Directed evolution of an enhanced POU reprogramming factor for cell fate engineering.

摘要: Transcription factor-driven cell fate engineering in pluripotency induction, transdifferentiation, and forward reprogramming require efficiency, speed, maturity for widespread adoption clinical translation. Here, we used Oct4, Sox2, Klf4, c-Myc driven to evaluate methods enhancing tailoring transitions, through directed evolution with iterative screening of pooled mutant libraries phenotypic selection. We identified an artificially evolved enhanced POU factor (ePOU) that substantially outperforms wild-type Oct4 terms speed efficiency. In contrast ePOU can induce Sox2 alone the absence three - ePOU/Klf4/c-Myc cocktails. Biochemical assays combined genome-wide analyses show acquires a new preference dimerize on palindromic DNA elements. Yet, moderate capacity function as pioneer factor, its bind octamer capability Sox17 proteins are not changed ePOU. Compared is thermodynamically stabilized persists longer cells. consequence, ePOU: (1) differentially activates several genes hitherto implicated reprogramming, (2) reveals unappreciated role thyrotropin-releasing hormone signaling, (3) binds distinct class retrotransposons. Collectively, these features enabled accelerate establishment network. This demonstrates selection novel variants from mammalian cells desired properties key advancing conversions biomolecules.