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Engineering modular and orthogonal genetic logic gates for robust digital-like synthetic biology

Author

Listed:
  • Baojun Wang

    (Imperial College London)

  • Richard I Kitney

    (Imperial College London)

  • Nicolas Joly

    (Faculty of Natural Sciences, Imperial College London
    Present address: Institut Jacques Monod, CNRS UMR 7592, Université Paris Diderot, Paris 75205, France.)

  • Martin Buck

    (Faculty of Natural Sciences, Imperial College London)

Abstract

Modular and orthogonal genetic logic gates are essential for building robust biologically based digital devices to customize cell signalling in synthetic biology. Here we constructed an orthogonal AND gate in Escherichia coli using a novel hetero-regulation module from Pseudomonas syringae. The device comprises two co-activating genes hrpR and hrpS controlled by separate promoter inputs, and a σ54-dependent hrpL promoter driving the output. The hrpL promoter is activated only when both genes are expressed, generating digital-like AND integration behaviour. The AND gate is demonstrated to be modular by applying new regulated promoters to the inputs, and connecting the output to a NOT gate module to produce a combinatorial NAND gate. The circuits were assembled using a parts-based engineering approach of quantitative characterization, modelling, followed by construction and testing. The results show that new genetic logic devices can be engineered predictably from novel native orthogonal biological control elements using quantitatively in-context characterized parts.

Suggested Citation

  • Baojun Wang & Richard I Kitney & Nicolas Joly & Martin Buck, 2011. "Engineering modular and orthogonal genetic logic gates for robust digital-like synthetic biology," Nature Communications, Nature, vol. 2(1), pages 1-9, September.
    • Handle: RePEc:nat:natcom:v:2:y:2011:i:1:d:10.1038_ncomms1516
    DOI: 10.1038/ncomms1516
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    Cited by:

    1. Kanakov, Oleg & Chen, Shangbin & Zaikin, Alexey, 2024. "Learning by selective plasmid loss for intracellular synthetic classifiers," Chaos, Solitons & Fractals, Elsevier, vol. 179(C).
    2. Trevor R. Simmons & Gina Partipilo & Ryan Buchser & Anna C. Stankes & Rashmi Srivastava & Darian Chiu & Benjamin K. Keitz & Lydia M. Contreras, 2024. "Rewiring native post-transcriptional global regulators to achieve designer, multi-layered genetic circuits," Nature Communications, Nature, vol. 15(1), pages 1-13, December.
    3. Weiyue Ji & Handuo Shi & Haoqian Zhang & Rui Sun & Jingyi Xi & Dingqiao Wen & Jingchen Feng & Yiwei Chen & Xiao Qin & Yanrong Ma & Wenhan Luo & Linna Deng & Hanchi Lin & Ruofan Yu & Qi Ouyang, 2013. "A Formalized Design Process for Bacterial Consortia That Perform Logic Computing," PLOS ONE, Public Library of Science, vol. 8(2), pages 1-9, February.
    4. Yuanli Gao & Lei Wang & Baojun Wang, 2023. "Customizing cellular signal processing by synthetic multi-level regulatory circuits," Nature Communications, Nature, vol. 14(1), pages 1-14, December.
    5. Lorenzo Pasotti & Nicolò Politi & Susanna Zucca & Maria Gabriella Cusella De Angelis & Paolo Magni, 2012. "Bottom-Up Engineering of Biological Systems through Standard Bricks: A Modularity Study on Basic Parts and Devices," PLOS ONE, Public Library of Science, vol. 7(7), pages 1-10, July.

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