A synthetic system for asymmetric cell division in Escherichia coli

Abstract

We describe a synthetic genetic circuit for controlling asymmetric cell division in Escherichia coli in which a progenitor cell creates a differentiated daughter cell while retaining its original phenotype. Specifically, we engineered an inducible system that can bind and segregate plasmid DNA to a single position in the cell. Upon cell division, colocalized plasmids are kept by one and only one of the daughter cells. The other daughter cell receives no plasmid DNA and is irreversibly differentiated from its sibling. In this way, we achieved asymmetric cell division through asymmetric plasmid partitioning. We then used this system to achieve physical separation of genetically distinct cells by tying motility to differentiation. Finally, we characterized an orthogonal inducible circuit that enables the simultaneous asymmetric partitioning of two plasmid species, resulting in cells that have four distinct differentiated states. These results point the way toward the engineering of multicellular systems from prokaryotic hosts.

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Fig. 1: Asymmetric plasmid partitioning in E. coli.
Fig. 2: Progenitor cells can undergo multiple rounds of asymmetric plasmid partitioning.
Fig. 3: Asymmetric plasmid partitioning with different origins of replication.
Fig. 4: Physical separation of genetically different cells.
Fig. 5: An orthogonal asymmetric plasmid partitioning system.

Data availability

The datasets generated and analyzed during the current study are available from the corresponding author upon reasonable request. All plasmids generated during this study are available on Addgene.

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Acknowledgements

We are especially grateful to L. Shapiro (Stanford) and C. Jacobs-Wagner (Yale) for providing plasmids and to K. Fahrner and H. Berg (Harvard) for her help in supplying and consulting on strain HCB84. This work was funded by the Defense Advanced Research Projects Agency Biological Technologies Biological Controls Program, award no. HR0011-17-2-0012 (approved for public release; distribution is unlimited) (M.R.B. and O.A.I.); the National Science Foundation through the joint NSF–National Institute of General Medical Sciences Mathematical Biology Program grant DMS-166290 (M.R.B.); the Welch Foundation grants C-1729 (M.R.B.) and C-1995 (O.A.I.); and the National Institutes of Health, grant R01GM117138 (M.R.B.).

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M.R.B., O.A.I. and S.M. conceived of the study. S.M., D.L.S. and S.P.B. performed experiments, and S.M. analyzed the data. M.R.B., O.A.I. and J.C. oversaw the project. All authors wrote the manuscript.

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Correspondence to Matthew R. Bennett.

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Molinari, S., Shis, D.L., Bhakta, S.P. et al. A synthetic system for asymmetric cell division in Escherichia coli. Nat Chem Biol 15, 917–924 (2019). https://doi.org/10.1038/s41589-019-0339-x

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