Compartmentalization is a defining feature of life that enables the spatial organization and maintenance of cellular biochemistry. Controlling spatial order holds promise for more efficient and programmable biomanufacturing applications and the construction of life-like systems from the bottom up. Biomolecular condensates represent an exciting engineering target in synthetic biology and cell-free biotechnology due to their capacity to self-assemble into dynamic compartments. Unlike membrane-bound organelles, biomolecular condensates allow molecular exchange with the surrounding medium. At the same time, they generate molecularly crowded niches that selectively recruit or exclude specific components, thereby creating unique environments. Their ability to rapidly respond to external cues by assembly or dissolution may enable control over biochemical processes, such as regulating gene expression, modulating enzymatic cascades, or orchestrating spatial organization in artificial cells and production systems.
Harnessing biomolecular condensates for programmable spatial organization in engineered systems, requires a thorough understanding of their assembly, internal dynamics and sequence-function relationships. Classical methods for studying condensates rely on reconstitution assays with purified proteins or genetic manipulation in living cells, each with major limitations. In cell-based studies, the high complexity of the cellular environment makes quantitative measurements challenging, and unknown molecular interactions often obscure functional insights. In vitro characterization benefits from defined reagent concentrations and conditions, but is hampered by laborious protein purification and the tendency of reactions to approach equilibrium, which does not reflect the non-equilibrium dynamics of cellular life.
The SYNSEMBL project addresses these gaps in current methods by developing a microfluidic, cell-free synthesis platform for characterization and engineering of biomolecular condensates under near-physiological, dynamically controlled conditions. The project aims to engineer condensate compartments with programmable properties, functions and to explore regulation strategies to dynamically control spatial organization. The expected impact includes accelerated design cycles for synthetic compartments, programmable, responsive materials, and advanced tools for sustainable biomanufacturing, biosensing, and understanding cellular organization.
