4.2 Article

A universal approach to gene expression engineering

Journal

SYNTHETIC BIOLOGY
Volume 7, Issue 1, Pages -

Publisher

OXFORD UNIV PRESS
DOI: 10.1093/synbio/ysac017

Keywords

artificial promoters; 5 ' untranslated region; transcription; translation

Funding

  1. Norwegian University of Science and Technology (NTNU)
  2. NTNU-Biotechnology, Enabling Technologies Program
  3. Faculty of Natural Sciences at NTNU
  4. Research Council of Norway [244278, 316129]
  5. EUH2020 MetaFluidics project [685474]

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In this study, a universal approach called Gene Expression Engineering (GeneEE) was developed to create artificial expression systems in various organisms, including bacteria and yeast, resulting in a wide range of expression levels.
In this study, we provide a universal approach to Gene Expression Engineering (GeneEE) for creating artificial expression systems. GeneEE leads to the generation of artificial 5 ' regulatory sequences (ARES) consisting of promoters and 5 ' untranslated regions. The ARES lead to the successful recruitment of RNA polymerase, related sigma factors and ribosomal proteins that result in a wide range of expression levels. We also demonstrate that by engaging native transcription regulators, GeneEE can be used to generate inducible promoters. To showcase the universality of the approach, we demonstrate that 200-nucleotide (nt)-long DNA with random composition can be used to generate functional expression systems in six bacterial species, Escherichia coli, Pseudomonas putida, Corynebacterium glutamicum, Thermus thermophilus, Streptomyces albus and Streptomyces lividans, and the eukaryote yeast Saccharomyces cerevisiae.

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