Synthetic Toolkit for Complex Genetic Circuit Engineering in Saccharomyces cerevisiae

Anssi Rantasalo, Joosu Kuivanen, Merja Penttilä, Jussi Jäntti, Dominik Mojzita* (Corresponding Author)

*Corresponding author for this work

    Research output: Contribution to journalArticleScientificpeer-review

    43 Citations (Scopus)

    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.

    Original languageEnglish
    Pages (from-to)1573-1587
    Number of pages15
    JournalACS Synthetic Biology
    Volume7
    Issue number6
    DOIs
    Publication statusPublished - 15 Jun 2018
    MoE publication typeA1 Journal article-refereed

    Funding

    This work was supported by the Finnish Funding Agency for Innovation (TEKES) (Living factories project). We thank Merja Helantera,̈ Oriane Vedrines, and Marjo Öster for technical assistance in the DNA cloning tasks, and Ulla Lahtinen for technical assistance in the UPLC analysis.

    Keywords

    • bistable switch
    • core promoter
    • gene regulation
    • synthetic transcription factor
    • yeast

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