A universal approach to gene expression engineering

Rahmi Lale; Lisa Tietze; Maxime Fages-Lartaud; Jenny Nesje; Ingerid Onsager; Kerstin Engelhardt; Che Fai Alex Wong; Madina Akan; Niklas Hummel; Jörn Kalinowski; Christian Rückert; Martin Frank Hohmann-Marriott

doi:10.1093/synbio/ysac017

A universal approach to gene expression engineering

Synth Biol (Oxf). 2022 Aug 22;7(1):ysac017. doi: 10.1093/synbio/ysac017. eCollection 2022.

Authors

Affiliations

¹ Department of Biotechnology, Faculty of Natural Sciences, Norwegian University of Science and Technology, Trondheim 7491, Norway.
² Department Microbiology and Biochemistry of Geisenheim University, Geisenheim 65366, Germany.
³ Feedstocks Division, Joint BioEnergy Institute, Emeryville 94608, CA, USA.
⁴ Department of Plant Biology, University of California, Davis 95616, CA, USA.
⁵ Center for Biotechnology, Bielefeld University, Bielefeld 33615, Germany.
⁶ United Scientists CORE (Limited), Dunedin 9014, New Zealand.

Abstract

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.

Keywords: 5′ untranslated region; artificial promoters; transcription; translation.