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A synthetic multicellular system for programmed pattern formation

Abstract

Pattern formation is a hallmark of coordinated cell behaviour in both single and multicellular organisms1,2,3. It typically involves cell–cell communication and intracellular signal processing. Here we show a synthetic multicellular system in which genetically engineered ‘receiver’ cells are programmed to form ring-like patterns of differentiation based on chemical gradients of an acyl-homoserine lactone (AHL) signal that is synthesized by ‘sender’ cells. In receiver cells, ‘band-detect’ gene networks respond to user-defined ranges of AHL concentrations. By fusing different fluorescent proteins as outputs of network variants, an initially undifferentiated ‘lawn’ of receivers is engineered to form a bullseye pattern around a sender colony. Other patterns, such as ellipses and clovers, are achieved by placing senders in different configurations. Experimental and theoretical analyses reveal which kinetic parameters most significantly affect ring development over time. Construction and study of such synthetic multicellular systems can improve our quantitative understanding of naturally occurring developmental processes and may foster applications in tissue engineering, biomaterial fabrication and biosensing.

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Figure 1: The band-detect multicellular system programs E. coli receiver cells to fluoresce only at intermediate distances from sender cells.
Figure 2: Simulated and experimental liquid-phase behaviour of high-detect and band-detect networks.
Figure 3: Experimental solid-phase behaviour of band-detect networks.
Figure 4: Ring formation dynamics.
Figure 5: Formation of various patterns.

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Acknowledgements

We thank D. Karig, S. Hooshangi, S. Thiberge, M.-T. Chen and S. Subramaniam for discussions or comments on the manuscript. This material is based on work supported by the Defense Advanced Research Projects Agency (DARPA).

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Correspondence to Ron Weiss.

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Supplementary Notes

This file contains Supplementary Table S1, which lists the high-detect, low-detect, sender, and marker plasmids used in this study. It also contains Supplementary Figure S1, which presents a contour map of band-detect gain as a function of LacI and CI repression efficiency; and Supplementary Figure S2, which details surface maps for BD2-Red and BD3 fluorescence intensities. (PDF 265 kb)

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Basu, S., Gerchman, Y., Collins, C. et al. A synthetic multicellular system for programmed pattern formation. Nature 434, 1130–1134 (2005). https://doi.org/10.1038/nature03461

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