Synthetic transcription amplifier system for orthogonal control of gene expression in saccharomyces cerevisiae

Anssi Rantasalo, Elena Czeizler, Riitta Virtanen, Juho Rousu, Harri Lähdesmäki, Merja Penttilä, Jussi Jäntti, Dominik Mojzita (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

13 Citations (Scopus)

Abstract

This work describes the development and characterization of a modular synthetic expression system that provides a broad range of adjustable and predictable expression levels in S. cerevisiae. The system works as a fixed-gain transcription amplifier, where the input signal is transferred via a synthetic transcription factor (sTF) onto a synthetic promoter, containing a defined core promoter, generating a transcription output signal. The system activation is based on the bacterial LexA-DNA-binding domain, a set of modified, modular LexA-binding sites and a selection of transcription activation domains. We show both experimentally and computationally that the tuning of the system is achieved through the selection of three separate modules, each of which enables an adjustable output signal: 1) the transcription-activation domain of the sTF, 2) the binding-site modules in the output promoter, and 3) the core promoter modules which define the transcription initiation site in the output promoter. The system has a novel bidirectional architecture that enables generation of compact, yet versatile expression modules for multiple genes with highly diversified expression levels ranging from negligible to very strong using one synthetic transcription factor. In contrast to most existing modular gene expression regulation systems, the present system is independent from externally added compounds. Furthermore, the established system was minimally affected by the several tested growth conditions. These features suggest that it can be highly useful in large scale biotechnology applications. © 2016 Rantasalo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Original languageEnglish
Article numbere0148320
JournalPLoS ONE
Volume11
Issue number2
DOIs
Publication statusPublished - 2016
MoE publication typeA1 Journal article-refereed

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Transcription
Gene expression
Yeast
Saccharomyces cerevisiae
Transcription Factors
transcription (genetics)
promoter regions
Gene Expression
gene expression
Transcriptional Activation
Chemical activation
Binding Sites
transcription factors
Gene expression regulation
transcriptional activation
Bacterial DNA
Gene Regulatory Networks
Transcription Initiation Site
Gene Expression Regulation
Licensure

Cite this

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title = "Synthetic transcription amplifier system for orthogonal control of gene expression in saccharomyces cerevisiae",
abstract = "This work describes the development and characterization of a modular synthetic expression system that provides a broad range of adjustable and predictable expression levels in S. cerevisiae. The system works as a fixed-gain transcription amplifier, where the input signal is transferred via a synthetic transcription factor (sTF) onto a synthetic promoter, containing a defined core promoter, generating a transcription output signal. The system activation is based on the bacterial LexA-DNA-binding domain, a set of modified, modular LexA-binding sites and a selection of transcription activation domains. We show both experimentally and computationally that the tuning of the system is achieved through the selection of three separate modules, each of which enables an adjustable output signal: 1) the transcription-activation domain of the sTF, 2) the binding-site modules in the output promoter, and 3) the core promoter modules which define the transcription initiation site in the output promoter. The system has a novel bidirectional architecture that enables generation of compact, yet versatile expression modules for multiple genes with highly diversified expression levels ranging from negligible to very strong using one synthetic transcription factor. In contrast to most existing modular gene expression regulation systems, the present system is independent from externally added compounds. Furthermore, the established system was minimally affected by the several tested growth conditions. These features suggest that it can be highly useful in large scale biotechnology applications. {\circledC} 2016 Rantasalo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",
author = "Anssi Rantasalo and Elena Czeizler and Riitta Virtanen and Juho Rousu and Harri L{\"a}hdesm{\"a}ki and Merja Penttil{\"a} and Jussi J{\"a}ntti and Dominik Mojzita",
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Synthetic transcription amplifier system for orthogonal control of gene expression in saccharomyces cerevisiae. / Rantasalo, Anssi; Czeizler, Elena; Virtanen, Riitta; Rousu, Juho; Lähdesmäki, Harri; Penttilä, Merja; Jäntti, Jussi; Mojzita, Dominik (Corresponding Author).

In: PLoS ONE, Vol. 11, No. 2, e0148320, 2016.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Rantasalo, Anssi

AU - Czeizler, Elena

AU - Virtanen, Riitta

AU - Rousu, Juho

AU - Lähdesmäki, Harri

AU - Penttilä, Merja

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AU - Mojzita, Dominik

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