Differentiation of Pluripotent Synthetic Minimal Cells via Genetic Circuits and Programmable Mating

Synthetic minimal cells, here defined as liposomal bioreactors synthesizing protein, are a recent technology that models the intricate gene and protein networks of live cells, without the noise inherent to natural systems. Here we show a toolset for engineering combinatorial genetic circuits in synthetic cells, where distinct populations of synthetic cells control single-gene components, and those populations are assembled via programmable fusion to construct complex biological pathways. We utilize this technology to demonstrate that progenitor populations can mimic differentiation into new lineages in response to small molecule stimuli or as an effect of “mating” with another synthetic cell population. This provides a practical tool for metabolic engineering and natural pathway study, as well as paving the way towards the construction of live synthetic cells with complex gene pathways and the ability to form distinct lineages.