Systematic Comparison and Rational Design of Theophylline Riboswitches for Effective Gene Repression
Xun Wang, Can Fang, Yifei Wang, Xinyu Shi, Fan Yu, Jin Xiong, Shan‐Ho Chou, Jin He
Abstract
A variety of gene expression regulation tools with significant regulatory effects are essential for the construction of complex gene circuits in synthetic biology. Riboswitches have received wide attention due to their unique biochemical, structural, and genetic properties. Here, we have not only systematically and precisely characterized the regulatory properties of previously developed theophylline riboswitches but also engineered a novel repressive theophylline riboswitch acting at the transcriptional level. By introducing coding sequences of a tandem riboswitch and a RepA protein degradation tag at the 5' end of the reporter gene, we successfully constructed a simple and effective regulatory cassette for gene regulation. Our work provides useful biological components for the construction of synthetic biology gene circuits.