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

Research output: Contribution to journalArticleResearchpeer-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 languageEnglish
Pages (from-to)3589
JournalNature Communications
Volume9
Issue number1
DOIs
Publication statusPublished - 4 Sep 2018
Externally publishedYes

Fingerprint

genes
Synthetic Biology
Libraries
Genes
Metabolic Networks and Pathways
Gene Expression
optimization
Metabolic engineering
Metabolic Engineering
biology
Pichia
Carotenoids
Yeast
Screening
Yeasts
carotene
yeast
pressing
Networks (circuits)
flexibility

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

Cite this

Engineered bidirectional promoters enable rapid multi-gene co-expression optimization. / Vogl, Thomas; Kickenweiz, Thomas; Pitzer, Julia; Sturmberger, Lukas; Weninger, Astrid; Biggs, Bradley W; Köhler, Eva-Maria; Baumschlager, Armin; Fischer, Jasmin Elgin; Hyden, Patrick; Wagner, Marlies; Baumann, Martina; Borth, Nicole; Geier, Martina; Ajikumar, Parayil Kumaran; Glieder, Anton.

In: Nature Communications, Vol. 9, No. 1, 04.09.2018, p. 3589.

Research output: Contribution to journalArticleResearchpeer-review

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
Vogl T, Kickenweiz T, Pitzer J, Sturmberger L, Weninger A, Biggs BW et al. Engineered bidirectional promoters enable rapid multi-gene co-expression optimization. Nature Communications. 2018 Sep 4;9(1):3589. https://doi.org/10.1038/s41467-018-05915-w
Vogl, Thomas ; Kickenweiz, Thomas ; Pitzer, Julia ; Sturmberger, Lukas ; Weninger, Astrid ; Biggs, Bradley W ; Köhler, Eva-Maria ; Baumschlager, Armin ; Fischer, Jasmin Elgin ; Hyden, Patrick ; Wagner, Marlies ; Baumann, Martina ; Borth, Nicole ; Geier, Martina ; Ajikumar, Parayil Kumaran ; Glieder, Anton. / Engineered bidirectional promoters enable rapid multi-gene co-expression optimization. In: Nature Communications. 2018 ; Vol. 9, No. 1. pp. 3589.
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