title: A portable expression resource for engineering cross-species genetic circuits and pathways.
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.
authors.affiliation: Department of Biological Engineering, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Manish
authors.identifier: 0000000288503272
authors.initials: M
authors.lastname: Kushwaha
authors.affiliation: 1] Department of Biological Engineering, Pennsylvania State University, University Park, Pennsylvania 16802, USA [2] Department of Chemical Engineering, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Howard M
authors.initials: HM
authors.lastname: Salis
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D053263:Gene Regulatory Networks; D005818:Genetic Engineering / Q000379:methods; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016958:Pseudomonas putida / Q000235:genetics*; D058615:Synthetic Biology / Q000379:methods*