Synthetic Promoters and Transcription Factors for Heterologous Protein Expression in Saccharomyces cerevisiae

Frontiers in Bioengineering and Biotechnology
Fabian MachensKatrin Messerschmidt

Abstract

Orthogonal systems for heterologous protein expression as well as for the engineering of synthetic gene regulatory circuits in hosts like Saccharomyces cerevisiae depend on synthetic transcription factors (synTFs) and corresponding cis-regulatory binding sites. We have constructed and characterized a set of synTFs based on either transcription activator-like effectors or CRISPR/Cas9, and corresponding small synthetic promoters (synPs) with minimal sequence identity to the host's endogenous promoters. The resulting collection of functional synTF/synP pairs confers very low background expression under uninduced conditions, while expression output upon induction of the various synTFs covers a wide range and reaches induction factors of up to 400. The broad spectrum of expression strengths that is achieved will be useful for various experimental setups, e.g., the transcriptional balancing of expression levels within heterologous pathways or the construction of artificial regulatory networks. Furthermore, our analyses reveal simple rules that enable the tuning of synTF expression output, thereby allowing easy modification of a given synTF/synP pair. This will make it easier for researchers to construct tailored transcriptional contr...Continue Reading

References

Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·M Gossen, H Bujard
Feb 1, 2002·Nature Biotechnology·Roger R Beerli, Carlos F Barbas
Apr 3, 2007·Nature Protocols·R Daniel Gietz, Robert H Schiestl
Jul 27, 2007·Proceedings of the National Academy of Sciences of the United States of America·Kevin F MurphyJames J Collins
Mar 17, 2009·Applied and Environmental Microbiology·Esben H HansenJørgen Hansen
Mar 26, 2009·Chemistry & Biology·Wilfried Weber, Martin Fussenegger
Apr 30, 2009·Annual Review of Phytopathology·Jens Boch, Ulla Bonas
Nov 26, 2009·Science·Matthew J Moscou, Adam J Bogdanove
Dec 17, 2009·Nature Biotechnology·Timothy K LuJames J Collins
Mar 19, 2011·Cell·Christina D Smolke, Pamela A Silver
Mar 23, 2011·Nucleic Acids Research·Robert MorbitzerThomas Lahaye
Jan 10, 2012·Nature Protocols·Neville E SanjanaFeng Zhang
Feb 18, 2012·Nature·Blake WiedenheftJennifer A Doudna
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Jul 12, 2012·Nature Biotechnology·Jana StreubelJens Boch
Feb 5, 2013·Nature Methods·Pablo Perez-PineraCharles A Gersbach
Feb 15, 2013·Nucleic Acids Research·Joshua F MecklerEnoch P Baldwin
Mar 6, 2013·Nucleic Acids Research·James E DiCarloGeorge M Church
Jul 31, 2013·Nature Methods·Morgan L MaederJ Keith Joung
Aug 28, 2013·ACS Synthetic Biology·Fahim FarzadfardTimothy K Lu
Jan 21, 2014·Nucleic Acids Research·Yanni LinGang Bao
Feb 27, 2014·Nucleic Acids Research·Alexandre JuilleratPhilippe Duchateau
Nov 26, 2014·Molecular Systems Biology·Alec A K Nielsen, Christopher A Voigt
Apr 8, 2015·The Plant Journal : for Cell and Molecular Biology·Kathleen BrücknerAlain Tissier
Sep 13, 2016·Scientific Reports·Sabine GogolokSteven M Pollard
Jan 29, 2017·Nucleic Acids Research·Lena HochreinBernd Mueller-Roeber
May 26, 2017·Nature Communications·Miles W GanderEric Klavins
Sep 16, 2017·Nucleic Acids Research·Lena HochreinBernd Mueller-Roeber

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Citations

Sep 27, 2018·Experimental Biology and Medicine·Jenny C Mortimer
May 18, 2020·Nature Communications·Gita Naseri, Mattheos A G Koffas
Feb 26, 2019·Molecular Biotechnology·Roghayyeh BaghbanMaryam Aria
Jan 23, 2021·Microbial Cell Factories·Michael Dare AsemoloyeLorenzo Pecoraro
Jul 17, 2021·Integrative Biology : Quantitative Biosciences From Nano to Macro·John McCarthy
May 12, 2018·ACS Synthetic Biology·Anssi RantasaloDominik Mojzita
Jan 8, 2021·ACS Synthetic Biology·James GilmanFilippo Menolascina
Aug 27, 2021·Frontiers in Bioengineering and Biotechnology·Gita NaseriChristoph Arenz

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Methods Mentioned

BETA
PCR
flow cytometry

Software Mentioned

BLAST
synTALE6
synTALEs
synTALE
TALE
SynTALE5
Flowing Software
AssemblX

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