In this paper, we present a strategy to integrate cell-free protein synthesis within agarose-based hydrogels that can be spatially organized and supplied by a synthetic “vasculature”. When supplying such vascularized hydrogels with reactants, larger components such as DNA plasmids are confined to the channels or immobilized in the gels while nanoscale reaction components can diffusively spread. We demonstrate spatial protein concentration profiles emerging from different cell-free gene circuits comprising production, gene activation, and negative feedback.
Bienau, A., Jäkel, A. C. & Simmel, F. C. Cell-Free Gene Expression in Bioprinted Fluidic Networks. ACS Synth. Biol. 13, 2447–2456 (2024).