Abstract

Synthetic biology has promoted the development of biosensors as tools for detecting trace substances. In the past, biosensors based on synthetic biology have been designed on living cells, but the development of cell biosensors has been greatly limited by defects such as the obstruction of cell membrane. However, the advent of cell-free synthetic biology addresses these limitations. Biosensors based on the cell-free protein synthesis system have the advantages of higher safety, higher sensitivity, and faster response time over cell biosensors, which makes cell-free biosensors have a broader application prospect. This review summarizes the workflow of various cell-free biosensors, including the identification of analytes and signal output. The detection range of cell-free biosensors is greatly enlarged by different recognition mechanisms and output methods. In addition, the review also discusses the applications of cell-free biosensors in environmental monitoring and health diagnosis, as well as existing deficiencies and aspects that should be improved. In the future, through continuous improvement and optimization, the potential of cell-free biosensors will be stimulated, and their application fields will be expanded.