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Iterative plug-and-play methodology for constructing and modifying synthetic gene networks

Nature Methods volume 9, pages 1077–1080 (2012)Cite this article

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Abstract

We present a methodology for the design, construction and modification of synthetic gene networks. This method emphasizes post-assembly modification of constructs based on network behavior, thus facilitating iterative design strategies and rapid tuning and repurposing of gene networks. The ease of post-construction modification afforded by this approach and the ever-increasing repository of components within the framework will help accelerate the development of functional genetic circuits for synthetic biology.

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Figure 1: Plug-and-play methodology for synthetic gene networks.
Figure 2: Construction and tuning of a bistable genetic toggle switch.
Figure 3: Transformation of the genetic toggle switch into functional three- and four-node feed-forward loops.

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Acknowledgements

This work was supported by the Howard Hughes Medical Institute and the Office of Naval Research Multidisciplinary University Research Initiative.

Author information

Author notes

  1. Kevin D Litcofsky and Raffi B Afeyan: These authors contributed equally to this work.

Authors and Affiliations

  1. Howard Hughes Medical Institute, Boston University, Boston, Massachusetts, USA

    Kevin D Litcofsky, Raffi B Afeyan, Russell J Krom, Ahmad S Khalil & James J Collins

  2. Department of Biomedical Engineering, Boston University, Boston, Massachusetts, USA

    Kevin D Litcofsky, Raffi B Afeyan, Russell J Krom, Ahmad S Khalil & James J Collins

  3. Center for BioDynamics, Boston University, Boston, Massachusetts, USA

    Kevin D Litcofsky, Raffi B Afeyan, Russell J Krom, Ahmad S Khalil & James J Collins

  4. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA

    James J Collins

Authors
  1. Kevin D Litcofsky
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  2. Raffi B Afeyan
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  3. Russell J Krom
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  4. Ahmad S Khalil
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  5. James J Collins
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Contributions

K.D.L., R.B.A., A.S.K. and J.J.C. designed the study. K.D.L., R.B.A., R.J.K. and A.S.K. performed experiments. K.D.L., R.B.A., R.J.K., A.S.K. and J.J.C. wrote the manuscript. A.S.K. and J.J.C. supervised the project.

Corresponding authors

Correspondence to Ahmad S Khalil or James J Collins.

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Competing interests

The authors declare no competing financial interests.

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Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Tables 1–3 and Supplementary Note (PDF 475 kb)

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Litcofsky, K., Afeyan, R., Krom, R. et al. Iterative plug-and-play methodology for constructing and modifying synthetic gene networks. Nat Methods 9, 1077–1080 (2012). https://doi.org/10.1038/nmeth.2205

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