Targeting riboswitches with synthetic small RNAs for metabolic engineering

Author(s)

Lins, Milca Rachel da Costa Ribeiro ^[1] ; Amorim, Laura Araujo da Silva ^[1] ; Correa, Graciely Gomes ^[1] ; Picao, Bruno Willian ^[1] ; Mack, Matthias ^[2] ; Cerri, Marcel Otavio ^[1] ; Pedrolli, Danielle Biscaro ^[1]

Affiliation

^[1] Univ Estadual Paulista UNESP, Sch Pharmaceut Sci, Dept Bioproc Engn & Biotechnol, Rodovia Araraquara Jau Km 1, BR-14800903 Araraquara, SP - Brazil

^[2] Mannheim Univ Appl Sci, Inst Tech Microbiol, Paul Wittsack Str 10, D-68163 Mannheim - Germany

• Document type: Journal article
• Source: METABOLIC ENGINEERING; v. 68, p. 59-67, NOV 2021.
• Web of Science Citations: 1

Abstract

Our growing knowledge of the diversity of non-coding RNAs in natural systems and our deepening knowledge of RNA folding and function have fomented the rational design of RNA regulators. Based on that knowledge, we designed and implemented a small RNA tool to target bacterial riboswitches and activate gene expression (rtRNA). The synthetic rtRNA is suitable for regulation of gene expression both in cell-free and in cellular systems. It targets riboswitches to promote the antitermination folding regardless the cognate metabolite concentration. Therefore, it prevents transcription termination increasing gene expression up to 103-fold. We successfully used small RNA arrays for multiplex targeting of riboswitches. Finally, we used the synthetic rtRNAs to engineer an improved riboflavin producer strain. The easiness to design and construct, and the fact that the rtRNA works as a single genome copy, make it an attractive tool for engineering industrial metabolite-producing strains. (AU)

• FAPESP's process: 20/08699-7 - Performance improvement in Airlift internal circulation bioreactor through geometric modifications to the inner tube
• Grantee: Marcel Otavio Cerri
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/17564-7 - Reprogramming the purine metabolism of Bacillus subtilis employing sRNA-technology
• Grantee: Danielle Biscaro Pedrolli
• Support Opportunities: Research Grants - Young Investigators Grants