Engineered bidirectional promoters enable rapid multi-gene co-expression optimization

Thomas Vogl; Thomas Kickenweiz; Julia Pitzer; Lukas Sturmberger; Astrid Weninger; Bradley W Biggs; Eva-Maria Köhler; Armin Baumschlager; Jasmin Elgin Fischer; Patrick Hyden; Marlies Wagner; Martina Baumann; Nicole Borth; Martina Geier; Parayil Kumaran Ajikumar; Anton Glieder

doi:10.1038/s41467-018-05915-w

Engineered bidirectional promoters enable rapid multi-gene co-expression optimization

Thomas Vogl, Thomas Kickenweiz, Julia Pitzer, Lukas Sturmberger, Astrid Weninger, Bradley W Biggs, Eva-Maria Köhler, Armin Baumschlager, Jasmin Elgin Fischer, Patrick Hyden, Marlies Wagner, Martina Baumann, Nicole Borth, Martina Geier, Parayil Kumaran Ajikumar, Anton Glieder

Institute of Molecular Biotechnology (6550)

Research output: Contribution to journal › Article › peer-review

Abstract

Numerous synthetic biology endeavors require well-tuned co-expression of functional components for success. Classically, monodirectional promoters (MDPs) have been used for such applications, but MDPs are limited in terms of multi-gene co-expression capabilities. Consequently, there is a pressing need for new tools with improved flexibility in terms of genetic circuit design, metabolic pathway assembly, and optimization. Here, motivated by nature's use of bidirectional promoters (BDPs) as a solution for efficient gene co-expression, we generate a library of 168 synthetic BDPs in the yeast Komagataella phaffii (syn. Pichia pastoris), leveraging naturally occurring BDPs as a parts repository. This library of synthetic BDPs allows for rapid screening of diverse expression profiles and ratios to optimize gene co-expression, including for metabolic pathways (taxadiene, β-carotene). The modular design strategies applied for creating the BDP library could be relevant in other eukaryotic hosts, enabling a myriad of metabolic engineering and synthetic biology applications.

Original language	English
Pages (from-to)	3589
Journal	Nature Communications
Volume	9
Issue number	1
DOIs	https://doi.org/10.1038/s41467-018-05915-w
Publication status	Published - 4 Sept 2018

Keywords

Promoter Regions, Genetic
Proteins
Diterpenes/metabolism
Farnesyltranstransferase/genetics
Gene Expression Regulation, Fungal
Genetic Engineering/methods
Histones/genetics
Microorganisms, Genetically-Modified
Pichia/genetics
beta Carotene/genetics

Fields of Expertise

Human- & Biotechnology

Access to Document

10.1038/s41467-018-05915-wLicence: CC BY 4.0

1 Finished
1 Active

A3b Industrial Enzymes by Recombinant DNA and Protein Engineering
Glieder, A. & Schwab, H.
1/01/95 → …
Project: Research project
Doctoral Program: Molecular Enzymology
Luley, C., Egger, S., Griengl, H., Lehsl, A., Lypetska, K., Malik, I. S., Schwab, H., Flis, V. V., Glieder, A., Fuchs, A., Marlingapla Halasiddappa, L., Hermetter, A., Pratter, S., Prattner, J., Freigaßner, M. E., Czabany, T., Schitter-Sollner, S., Macheroux, P., Eixelsberger, T., Thonhauser, M., Horvath, S., Nidetzky, B., Winkler, A., Daum, G., Connerth, M. & Rajakumari, S.
1/01/05 → 31/12/17
Project: Research project

Cite this

Vogl, T., Kickenweiz, T., Pitzer, J., Sturmberger, L., Weninger, A., Biggs, B. W., Köhler, E-M., Baumschlager, A., Fischer, J. E., Hyden, P., Wagner, M., Baumann, M., Borth, N., Geier, M., Ajikumar, P. K., & Glieder, A. (2018). Engineered bidirectional promoters enable rapid multi-gene co-expression optimization. Nature Communications , 9(1), 3589. https://doi.org/10.1038/s41467-018-05915-w

Vogl, T, Kickenweiz, T, Pitzer, J, Sturmberger, L, Weninger, A, Biggs, BW, Köhler, E-M, Baumschlager, A, Fischer, JE, Hyden, P, Wagner, M, Baumann, M, Borth, N, Geier, M, Ajikumar, PK & Glieder, A 2018, 'Engineered bidirectional promoters enable rapid multi-gene co-expression optimization', Nature Communications , vol. 9, no. 1, pp. 3589. https://doi.org/10.1038/s41467-018-05915-w

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abstract = "Numerous synthetic biology endeavors require well-tuned co-expression of functional components for success. Classically, monodirectional promoters (MDPs) have been used for such applications, but MDPs are limited in terms of multi-gene co-expression capabilities. Consequently, there is a pressing need for new tools with improved flexibility in terms of genetic circuit design, metabolic pathway assembly, and optimization. Here, motivated by nature's use of bidirectional promoters (BDPs) as a solution for efficient gene co-expression, we generate a library of 168 synthetic BDPs in the yeast Komagataella phaffii (syn. Pichia pastoris), leveraging naturally occurring BDPs as a parts repository. This library of synthetic BDPs allows for rapid screening of diverse expression profiles and ratios to optimize gene co-expression, including for metabolic pathways (taxadiene, β-carotene). The modular design strategies applied for creating the BDP library could be relevant in other eukaryotic hosts, enabling a myriad of metabolic engineering and synthetic biology applications.",

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KW - Farnesyltranstransferase/genetics

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KW - Genetic Engineering/methods

KW - Histones/genetics

KW - Microorganisms, Genetically-Modified

KW - Pichia/genetics

KW - beta Carotene/genetics

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Engineered bidirectional promoters enable rapid multi-gene co-expression optimization

Abstract

Keywords

Fields of Expertise

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Projects

A3b Industrial Enzymes by Recombinant DNA and Protein Engineering

Doctoral Program: Molecular Enzymology

Cite this