title: Automated optogenetic feedback control for precise and robust regulation of gene expression and cell growth.
abstract: Dynamic control of gene expression can have far-reaching implications for biotechnological applications and biological discovery. Thanks to the advantages of light, optogenetics has emerged as an ideal technology for this task. Current state-of-the-art methods for optical expression control fail to combine precision with repeatability and cannot withstand changing operating culture conditions. Here, we present a novel fully automatic experimental platform for the robust and precise long-term optogenetic regulation of protein production in liquid Escherichia coli cultures. Using a computer-controlled light-responsive two-component system, we accurately track prescribed dynamic green fluorescent protein expression profiles through the application of feedback control, and show that the system adapts to global perturbations such as nutrient and temperature changes. We demonstrate the efficacy and potential utility of our approach by placing a key metabolic enzyme under optogenetic control, thus enabling dynamic regulation of the culture growth rate with potential applications in bacterial physiology studies and biotechnology.
authors: Department of Biosystems Science and Engineering, ETH Zurich, Basel 4058, Switzerland.
authors: Andreas
authors: A
authors: Milias-Argeitis
authors: Department of Biosystems Science and Engineering, ETH Zurich, Basel 4058, Switzerland.
authors: Marc
authors: M
authors: Rullan
authors: Department of Biosystems Science and Engineering, ETH Zurich, Basel 4058, Switzerland.
authors: Stephanie K
authors: SK
authors: Aoki
authors: Department of Biosystems Science and Engineering, ETH Zurich, Basel 4058, Switzerland.
authors: Peter
authors: P
authors: Buchmann
authors: Department of Biosystems Science and Engineering, ETH Zurich, Basel 4058, Switzerland.
authors: Mustafa
authors: M
authors: Khammash
mesh_terms: D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002453:Cell Cycle; D049109:Cell Proliferation; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005246:Feedback; D005786:Gene Expression Regulation; D008715:Methionine / Q000096:biosynthesis; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism; D062308:Optogenetics / Q000295:instrumentation / Q000379:methods*
chemical_list: D029968:Escherichia coli Proteins; D008715:Methionine; D008780:Methyltransferases; C058170:metE protein, E coli