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Artificial Transcription Factors for Tuneable Gene Expression in . | LitMetric

The non-conventional yeast (syn. ) has become a powerful eukaryotic expression platform for biopharmaceutical and biotechnological applications on both laboratory and industrial scales. Despite the fundamental role that artificial transcription factors (ATFs) play in the orthogonal control of gene expression in synthetic biology, a limited number of ATFs are available for . To establish orthogonal regulators for use in , we characterized ATFs derived from Arabidopsis TFs. The plant-derived ATFs contain the binding domain of TFs from the plant , in combination with the activation domains of yeast and plant and a synthetic promoter harboring the cognate -regulatory motifs. Chromosomally integrated ATFs and their binding sites (ATF/BSs) resulted in a wide spectrum of inducible transcriptional outputs in , ranging from as low as 1- to as high as ∼63-fold induction with only small growth defects. We demonstrated the application of ATF/BSs by generating cells that produce β-carotene. Notably, the productivity of β-carotene in was ∼4.8-fold higher than that in , reaching ∼59% of the β-carotene productivity obtained in a strain optimized for the production of the β-carotene precursor, farnesyl diphosphate, by rewiring the endogenous metabolic pathways using plant-derived ATF/BSs. Our data suggest that plant-derived regulators have a high degree of transferability from to . The plant-derived ATFs, together with their cognate binding sites, powerfully increase the repertoire of transcriptional regulatory modules for the tuning of protein expression levels required in metabolic engineering or synthetic biology in .

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8381338PMC
http://dx.doi.org/10.3389/fbioe.2021.676900DOI Listing

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