Multicellularity enables the growth of complex life forms as it allows for the specialization of cell types, differentiation and large-scale spatial organization. In a similar way, modular construction of synthetic multicellular systems will lead to dynamic biomimetic materials that can respond to their environment in complex ways. To achieve this goal, artificial cellular communication and developmental programs still have to be established. Here, we create geometrically controlled spatial arrangements of emulsion-based artificial cellular compartments containing synthetic in vitro gene circuitry, separated by lipid bilayer membranes. We quantitatively determine the membrane pore-dependent response of the circuits to artificial morphogen gradients, which are established via diffusion from dedicated organizer cells. Utilizing different types of feedforward and feedback in vitro gene circuits, we then implement artificial signalling and differentiation processes, demonstrating the potential for the realization of complex spatiotemporal dynamics in artificial multicellular systems.
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Raw data used for the generation of the figures are available from the authors upon request. Plasmids pSB1A3-AD009, pSB1A3-AD010 and pSB1A3-AD011 are available on Addgene.
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This work was supported by the European Research Council (grant agreement no. 694410, AEDNA) and the DFG Cluster of Excellence Nanosystems Initiative Munich (DFG EXC 4/3). A.D. acknowledges additional support by the DFG Research Training Group ‘Chemical Foundations of Synthetic Biology’ GRK 2062/1. The authors thank B. Tinao for her preliminary work on the artificial multicellular assemblies. They also thank D. Ziegler and J. List for their help with set-up construction, S. Sagredo and E. Falgenhauer for providing purified enzymes, and M. Schwarz-Schilling for helpful discussions. Correspondence and request for materials should be addressed to F.C.S.
The authors declare no competing interests.
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Supplementary methods, text, figures, tables, videos and references
This video shows the time lapse of Fig. 1b–c
Time lapse of the diffusion of DFHBI into a network of receivers containing a Spinach RNA transcription
This video shows 6 arrays containing the pulse propagation circuit, with different numbers of receivers
These videos show 2D networks containing the pulse generator
This video shows specifically a time-lapse of Fig. 3g
This video shows a differentiating network of one sender (indicated by S) and 4 receivers containing the circuit described in Fig. 4b, left panel
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Dupin, A., Simmel, F.C. Signalling and differentiation in emulsion-based multi-compartmentalized in vitro gene circuits. Nature Chem 11, 32–39 (2019). https://doi.org/10.1038/s41557-018-0174-9
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