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A portable expression resource for engineering cross-species genetic circuits and pathways

Author

Listed:
  • Manish Kushwaha

    (Pennsylvania State University)

  • Howard M. Salis

    (Pennsylvania State University
    Pennsylvania State University)

Abstract

Genetic circuits and metabolic pathways can be reengineered to allow organisms to process signals and manufacture useful chemicals. However, their functions currently rely on organism-specific regulatory parts, fragmenting synthetic biology and metabolic engineering into host-specific domains. To unify efforts, here we have engineered a cross-species expression resource that enables circuits and pathways to reuse the same genetic parts, while functioning similarly across diverse organisms. Our engineered system combines mixed feedback control loops and cross-species translation signals to autonomously self-regulate expression of an orthogonal polymerase without host-specific promoters, achieving nontoxic and tuneable gene expression in diverse Gram-positive and Gram-negative bacteria. Combining 50 characterized system variants with mechanistic modelling, we show how the cross-species expression resource’s dynamics, capacity and toxicity are controlled by the control loops’ architecture and feedback strengths. We also demonstrate one application of the resource by reusing the same genetic parts to express a biosynthesis pathway in both model and non-model hosts.

Suggested Citation

  • Manish Kushwaha & Howard M. Salis, 2015. "A portable expression resource for engineering cross-species genetic circuits and pathways," Nature Communications, Nature, vol. 6(1), pages 1-11, November.
  • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8832
    DOI: 10.1038/ncomms8832
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    Cited by:

    1. Amir Pandi & Christoph Diehl & Ali Yazdizadeh Kharrazi & Scott A. Scholz & Elizaveta Bobkova & Léon Faure & Maren Nattermann & David Adam & Nils Chapin & Yeganeh Foroughijabbari & Charles Moritz & Nic, 2022. "A versatile active learning workflow for optimization of genetic and metabolic networks," Nature Communications, Nature, vol. 13(1), pages 1-15, December.
    2. Baiyang Liu & Christian Cuba Samaniego & Matthew R. Bennett & Elisa Franco & James Chappell, 2023. "A portable regulatory RNA array design enables tunable and complex regulation across diverse bacteria," Nature Communications, Nature, vol. 14(1), pages 1-10, December.
    3. Lauren Gambill & August Staubus & Kim Wai Mo & Andrea Ameruoso & James Chappell, 2023. "A split ribozyme that links detection of a native RNA to orthogonal protein outputs," Nature Communications, Nature, vol. 14(1), pages 1-12, December.

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