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


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

Author information




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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The authors declare no competing financial interests.

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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).

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