Design of orthogonal regulatory systems for modulating gene expression in plants

• Published in: Nature chemical biology Vol. 16; no. 8; pp. 857 - 865
• Main Authors: Belcher, Michael S., Vuu, Khanh M., Zhou, Andy, Mansoori, Nasim, Agosto Ramos, Amanda, Thompson, Mitchell G., Scheller, Henrik V., Loqué, Dominique, Shih, Patrick M.
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.08.2020; Nature Publishing Group
• Subjects: 631/337/572; 631/449; 631/92/552; Agricultural biotechnology; Agricultural practices; Arabidopsis - genetics; Arabidopsis thaliana; Biochemical Engineering; Biochemistry; Bioorganic Chemistry; Biotechnology; Cell Biology; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Deoxyribonucleic acid; Design; Design factors; DNA; Environmental effects; Gene expression; Gene Expression - genetics; Gene Expression Regulation, Plant - genetics; Gene Expression Regulation, Plant - physiology; Gene regulation; Gene Regulatory Networks - genetics; Genes; Nicotiana - genetics; Promoter Regions, Genetic - genetics; Regulatory Elements, Transcriptional - genetics; Repressors; Saccharomyces - enzymology; Saccharomyces - genetics; Toolkits; Transcription factors; Transcription Factors - metabolism; Transgenes; Yeasts
• ISSN: 1552-4450; 1552-4469
• DOI: 10.1038/s41589-020-0547-4

Agricultural biotechnology strategies often require the precise regulation of multiple genes to effectively modify complex plant traits. However, most efforts are hindered by a lack of characterized tools that allow for reliable and targeted expression of transgenes. We have successfully engineered a library of synthetic transcriptional regulators that modulate expression strength in planta. By leveraging orthogonal regulatory systems from Saccharomyces spp., we have developed a strategy for the design of synthetic activators, synthetic repressors, and synthetic promoters and have validated their use in Nicotiana benthamiana and Arabidopsis thaliana . This characterization of contributing genetic elements that dictate gene expression represents a foundation for the rational design of refined synthetic regulators. Our findings demonstrate that these tools provide variation in transcriptional output while enabling the concerted expression of multiple genes in a tissue-specific and environmentally responsive manner, providing a basis for generating complex genetic circuits that process endogenous and environmental stimuli. Engineering of yeast transcription factors and design of hybrid DNA promoter elements have resulted in a toolkit for tunable and orthogonal regulation of gene expression in Arabidopsis thaliana and Nicotiana benthamiana plants.