Construction of a genetic toggle switch in Escherichia coli (original) (raw)

Nature volume 403, pages 339–342 (2000)Cite this article

Abstract

It has been proposed1 that gene-regulatory circuits with virtually any desired property can be constructed from networks of simple regulatory elements. These properties, which include multistability and oscillations, have been found in specialized gene circuits such as the bacteriophage λ switch2 and the Cyanobacteria circadian oscillator3. However, these behaviours have not been demonstrated in networks of non-specialized regulatory components. Here we present the construction of a genetic toggle switch—a synthetic, bistable gene-regulatory network—in Escherichia coli and provide a simple theory that predicts the conditions necessary for bistability. The toggle is constructed from any two repressible promoters arranged in a mutually inhibitory network. It is flipped between stable states using transient chemical or thermal induction and exhibits a nearly ideal switching threshold. As a practical device, the toggle switch forms a synthetic, addressable cellular memory unit and has implications for biotechnology, biocomputing and gene therapy.

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References

  1. Modod, J. & Jacob, F. General conclusions: teleonomic mechanisms in cellular metabolism, growth and differentiation. Cold Spring Harb. Symp. Quant. Biol. 26, 389–401 (1961).
    Article Google Scholar
  2. Ptashne, M. A Genetic Switch: Phage λ and Higher Organisms (Cell, Cambridge, Massachusetts, 1992).
    Google Scholar
  3. Ishiura, M. et al. Expression of a gene cluster kaiABC as a circadian feedback process in cyanobacteria. Science 281, 1519–1523 (1998).
    Article ADS CAS PubMed Google Scholar
  4. Schellenberger, W., Eschrich, K. & Hofmann, E. Self-organization of a glycolytic reconstituted enzyme system: alternate stable stationary states, hysteretic transitions and stabilization of the energy charge. Adv. Enzyme Regul. 19, 257–284 (1980).
    Article CAS PubMed Google Scholar
  5. Glass, L. & Kauffman, S. A. The logical analysis of continuous, non-linear biochemical control networks. J. Theor. Biol. 39, 103–129 (1973).
    Article CAS PubMed Google Scholar
  6. Glass, L. Classification of biological networks by their qualitative dynamics. J. Theor. Biol. 54, 85–107 (1975).
    Article CAS PubMed Google Scholar
  7. Glass, L. Combinatorial and topological methods in nonlinear chemical kinetics. J. Chem. Phys. 63, 1325–1335 (1975).
    Article ADS CAS Google Scholar
  8. Kauffman, S. The large scale structure and dynamics of gene control circuits: an ensemble approach. J. Theor. Biol. 44, 167– 190 (1974).
    Article CAS PubMed Google Scholar
  9. Thomas, R. Logical analysis of systems comprising feedback loops. J. Theor. Biol. 73, 631–656 ( 1978).
    Article CAS PubMed Google Scholar
  10. Thomas, R. Regulatory networks seen as asynchronous automata: a logical description. J. Theor. Biol. 153, 1– 23 (1991).
    Article Google Scholar
  11. Tchuraev, R. N. A new method for the analysis of the dynamics of the molecular genetic control systems. I. Description of the method of generalized threshold models. J. Theor. Biol. 151, 71–87 (1991).
    Article CAS PubMed Google Scholar
  12. Arkin, A. & Ross, J. Computational functions in biochemical reaction networks. Biophys. J. 67, 560– 578 (1994).
    Article ADS CAS PubMed PubMed Central Google Scholar
  13. Bhalla, U. S. & Iyengar, R. Emergent properties of networks of biological signaling pathways. Science 283, 381–387 (1999).
    Article ADS CAS PubMed Google Scholar
  14. Yagilo, G. & Yagil, E. On the relation between effector concentration and the rate of induced enzyme synthesis. Biophys. J. 11, 11–27 (1971).
    Article ADS Google Scholar
  15. Shea, M. A. & Ackers, G. K. The OR control system of bacteriophage Lambda: a physical-chemical model for gene regulation. J. Mol. Biol. 181, 211–230 (1985).
    Article CAS PubMed Google Scholar
  16. Smith, T. F., Sadler, J. R. & Goad, W. Statistical–mechanical modeling of a regulatory protein: the Lactose repressor. Math. Biosci. 36, 61–86 (1977).
    Article CAS Google Scholar
  17. Arkin, A., Ross, J. & McAdams, H. H. Stochastic kinetic analysis of developmental pathway bifurcation in phage λ-infected Escherichia coli cells. Genetics 149, 1633–1648 (1998).
    CAS PubMed PubMed Central Google Scholar
  18. McAdams, H. H. & Arkin, A. Stochastic mechanisms in gene expression. Proc. Natl Acad. Sci. USA 94, 814–819 (1997).
    Article ADS CAS PubMed PubMed Central Google Scholar
  19. McAdams, H. H. & Arkin, A. Stimulation of prokaryotic genetic circuits. Annu. Rev. Biophys. Biomol. Struct. 27, 199–224 (1998).
    Article CAS PubMed Google Scholar
  20. Lutz, R. & Bujard, H. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 25, 1203–1210 (1997).
    Article CAS PubMed PubMed Central Google Scholar
  21. Cormack, B. P., Valdivia, R. H. & Falkow, S. FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173, 33–38 (1996).
    Article CAS PubMed Google Scholar
  22. Ausubel, F. M. et al. Current Protocols in Molecular Biology (Wiley, New York, 1987).
    Google Scholar
  23. Sambrook, J., Fritsch, E. F. & Maniatis, T. Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Plainview, New York, 1989).
    Google Scholar
  24. Edelstein-Keshet, L. Mathematical Models in Biology (McGraw-Hill, New York, 1988).
    MATH Google Scholar
  25. Kaplan, D. & Glass, L. Understanding Nonlinear Dynamics (Springer, New York, 1995).
    Book Google Scholar
  26. Yagil, E. & Yagil, G. On the relation between effector concentration and the rate of induced enzyme synthesis. Biophys. J. 11, 11–27 (1971).
    Article ADS CAS PubMed PubMed Central Google Scholar
  27. Rubinow, S. I. Introduction to Mathematical Biology (Wiley, New York, 1975).
    MATH Google Scholar

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Acknowledgements

We thank M. Bitensky and T. Yoshida for providing access to their flow cytometer; Y. Yu for his suggestions on plasmid construction; C. Sabanayagam for his technical advice; and C. Chow for his mathematical advice. This work was supported by the Office of Naval Research and the College of Engineering at Boston University.

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Authors and Affiliations

  1. Department of Biomedical Engineering,
    Timothy S. Gardner, Charles R. Cantor & James J. Collins
  2. Center for BioDynamics,
    Timothy S. Gardner & James J. Collins

Authors

  1. Timothy S. Gardner
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  2. Charles R. Cantor
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  3. James J. Collins
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Corresponding author

Correspondence toJames J. Collins.

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Center for Advanced Biotechnology, Boston University, 44 Cummington Street, Boston, Massachusetts 02215, USA

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Gardner, T., Cantor, C. & Collins, J. Construction of a genetic toggle switch in Escherichia coli.Nature 403, 339–342 (2000). https://doi.org/10.1038/35002131

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