Targeting riboswitches with synthetic small RNAs for metabolic engineering

Lins, Milca Rachel da Costa Ribeiro [UNESP]; Amorim, Laura Araujo da Silva [UNESP]; Corrêa, Graciely Gomes [UNESP]; Picão, Bruno Willian [UNESP]; Mack, Matthias; Cerri, Marcel Otávio [UNESP]; Pedrolli, Danielle Biscaro [UNESP]

Targeting riboswitches with synthetic small RNAs for metabolic engineering

Data

2021-11-01

Autores

Lins, Milca Rachel da Costa Ribeiro

Amorim, Laura Araujo da Silva

Corrêa, Graciely Gomes

Picão, Bruno Willian

Mack, Matthias

Cerri, Marcel Otávio

Pedrolli, Danielle Biscaro

Tipo

Artigo

Resumo

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.