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

Manish Kushwaha, Howard M. Salis

Research output: Contribution to journalArticle

54 Citations (Scopus)

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.

Original languageEnglish (US)
Article number7832
JournalNature communications
Volume6
DOIs
StatePublished - Jul 17 2015

Fingerprint

Genetic Crosses
organisms
resources
engineering
Synthetic Biology
Metabolic engineering
Metabolic Engineering
Networks (circuits)
Biosynthesis
Metabolic Networks and Pathways
Gram-Negative Bacteria
Gene expression
Feedback control
Toxicity
Bacteria
biosynthesis
reuse
gene expression
feedback control
Feedback

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

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A portable expression resource for engineering cross-species genetic circuits and pathways. / Kushwaha, Manish; Salis, Howard M.

In: Nature communications, Vol. 6, 7832, 17.07.2015.

Research output: Contribution to journalArticle

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