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.

我们对自然系统中非编码 RNA 多样性的日益了解以及对 RNA 折叠和功能的深入了解促进了 RNA 调节剂的合理设计。基于这些知识，我们设计并实施了一种小 RNA 工具来靶向细菌核糖开关并激活基因表达 (rtRNA)。合成的 rtRNA 适用于在无细胞和细胞系统中调节基因表达。无论同源代谢物浓度如何，它都以核糖开关为目标，以促进抗终止折叠。因此，它可以防止转录终止，使基因表达增加 103 倍。我们成功地将小 RNA 阵列用于核糖开关的多重靶向。最后，我们使用合成的 rtRNA 来设计改进的核黄素生产菌株。易于设计和建造，