Reconstructing cellular complexity is a central challenge in synthetic biology, with profound implications for understanding life and advancing bio-inspired nanotechnologies. A critical step toward this goal is replicating the dynamic interplay among membrane components and their functions. Here, we demonstrate a double-necked synthetic cell microreactor (DCM) that incorporates two dynamic, DNA-based pores in the membrane of a giant unilamellar vesicle (GUV). The formation of the DCM leverages a signaling pathway mediated by GUV membrane dynamics to coordinate interactions between light-responsive small pores and self-arranged sealable large pores. This system enables sequential, on-demand delivery of molecular reactants with high spatiotemporal precision. Using DCMs, we demonstrate confined biochemical reactions, including a glucose oxidase-myoglobin cascade, cytoskeleton-mimetic actin polymerization and bundling, cell-free Spinach RNA transcription, and the synthesis of threedimensional DNA crystals that extend beyond natural systems. By coupling orchestrated multistep signaling with dynamic control of membrane permeability, the DCM establishes a versatile platform for emulating and expanding the functional complexity of natural cellular systems.
For reference, see: A synthetic cell microreactor with two types of interacting dynamic DNA-based pores. Fan, S., Ding, L.J., Renz, B., Liu A.P., Speck, T., Yan, H., Nussberger, S. & Liu, N. Nature Chemistry, doi:10.1038/s41557-026-02124-7 (2026)