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 journalArticlepeer-review


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
Issue number1
Publication statusPublished - 4 Sept 2018


  • 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

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