High content design of riboswitch biosensors: All-around rational module-by-module design

Riboswitches are ideal, diverse regulatory elements with simple mechanisms. Therefore, they are widely employed in synthetic biology to construct highly efficient biosensors for biosensing and bioimaging. This review presents a rational riboswitch biosensor design that combines computational and hybrid methods. Four main steps are discussed, including the rational design of riboswitch screening libraries, rational riboswitch screening, rational tailoring, and the rational selection of reporting elements. Furthermore, the application of riboswitch biosensors for molecular detection and biological imaging is introduced. Finally, the future development and construction of ideal riboswitch biosensors are discussed, while assessing their application for point-of-care testing, health monitoring, and gene therapy. This review is expected to provide a comprehensive overview of the rational construction of riboswitch biosensors.