Synthetic Toolkit for Complex Genetic Circuit Engineering in Saccharomyces cerevisiae

ACS Synthetic Biology
Anssi RantasaloDominik Mojzita

Abstract

Sustainable production of chemicals, materials, and pharmaceuticals is increasingly performed by genetically engineered cell factories. Engineering of complex metabolic routes or cell behavior control systems requires robust and predictable gene expression tools. In this challenging task, orthogonality is a fundamental prerequisite for such tools. In this study, we developed and characterized in depth a comprehensive gene expression toolkit that allows accurate control of gene expression in Saccharomyces cerevisiae without marked interference with native cellular regulation. The toolkit comprises a set of transcription factors, designed to function as synthetic activators or repressors, and transcription-factor-dependent promoters, which together provide a broad expression range surpassing, at high end, the strongest native promoters. Modularity of the developed tools is demonstrated by establishing a novel bistable genetic circuit with robust performance to control a heterologous metabolic pathway and enabling on-demand switching between two alternative metabolic branches.

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Citations

Jul 28, 2019·Applied Microbiology and Biotechnology·Kristoffer KrogerusBrian Gibson
May 2, 2020·Nature Communications·Benjamin J Kotopka, Christina D Smolke
Apr 25, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Attila Becskei
Dec 31, 2019·Molecular Systems Biology·Francisca MonteiroMatthias Heinemann
Feb 5, 2021·Microbial Cell Factories·Joosu KuivanenJussi Jäntti
Apr 18, 2021·FEMS Yeast Research·Louis C Dacquay, David R McMillen
May 6, 2019·ACS Synthetic Biology·Arun S RajkumarJay D Keasling
Sep 4, 2021·FEMS Yeast Research·Maurizio MorminoYvonne Nygård
Oct 14, 2021·Applied Microbiology and Biotechnology·Kristoffer KrogerusRichard Preiss
Nov 9, 2021·Molecular Systems Biology·Daniel A Anderson, Christopher A Voigt
Sep 30, 2021·Journal of Microbiology and Biotechnology·Aqsa AhmedZhonghu Bai

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Methods Mentioned

BETA
flow cytometry
PCR
flow

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