Synthetic Transcription Amplifier System for Orthogonal Control of Gene Expression in Saccharomyces cerevisiae

PloS One
Anssi RantasaloDominik Mojzita

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 tra...Continue Reading

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Citations

Apr 28, 2017·Journal of Experimental Botany·Tessa MosesAlain Goossens
Nov 30, 2018·Molecular Systems Biology·Felix MoserChristopher A Voigt
Nov 20, 2019·Microbial Cell Factories·László MózsikYvonne Nygård
Jul 25, 2018·Current Protocols in Protein Science·Helena NevalainenNatalie Curach
Nov 4, 2017·Frontiers in Bioengineering and Biotechnology·Fabian MachensKatrin Messerschmidt
Feb 26, 2019·Molecular Biotechnology·Roghayyeh BaghbanMaryam Aria
Aug 5, 2020·Nature Microbiology·Ye ChenChristopher A Voigt
May 12, 2018·ACS Synthetic Biology·Anssi RantasaloDominik Mojzita
Aug 6, 2021·Biotechnology Journal·Andrew P Cazier, John Blazeck
Sep 4, 2021·FEMS Yeast Research·Maurizio MorminoYvonne Nygård

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Datasets Mentioned

BETA
MODEL1510230005

Methods Mentioned

BETA
electrophoretic mobility shift assay
PCR
electrophoresis

Software Mentioned

Advance Relative Quantification
pBID2
ED
COPASI

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