Rational design and optimization of synthetic gene switches for controlling cell-fate decisions in pluripotent stem cells

Advances in synthetic biology have enabled robust control of cell behavior by using tunable genetic circuits to regulate gene expression a ligand-dependent manner. Such can be used direct the differentiation pluripotent stem cells (PSCs) towards desired types, but rational design PSCs is challenging due variable intracellular environment. Here, we provide framework for implementing switches based on combinations transcriptional, structural, and posttranslational elements that engineered as required, vanillic acid-controlled transcriptional activator (VanA) model system. We further show VanA system multiplexed with well-established reverse tetracycline-controlled (rtTA) enable independent different transcription factors human induced order enhance lineage specification early pancreatic progenitors. This work represents first step standardizing construction building gene-regulatory networks guide fate.