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Construction of a genetic toggle switch in Escherichia coli

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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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Figure 1: Toggle switch design.
Figure 2: Geometric structure of the toggle equations.
Figure 3: The toggle switch plasmid.
Figure 4: Demonstration of bistability.
Figure 5: Toggle switch induction threshold.
Figure 6: TAK117 switching time.

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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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Correspondence to James 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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