Application of Sensor Based on Circularly Permuted Fluorescent Protein in Molecular Biology, Metabolism, and Cellular Imaging

ABSTRACT Circular permutation is a natural phenomenon involving amino acid sequence rearrangement during protein evolution. This approach relies on the native protein sequence and reorganizes the connectivity of secondary structural elements by rearranging the amino acid sequence, without altering the overall protein structure. Circularly permuted fluorescent proteins (cpFPs) have received widespread attention in recent years for their applications in the field of sensors. Rearranging the structure of fluorescent proteins in a circular permutation, the relative positions of their termini can be effectively altered, thereby adjusting their sensitivity to environmental signals such as pH, ion concentration, voltage, or molecular interactions. This property has been widely used to monitor intracellular processes dynamically, particularly in studying neural activity, metabolic reactions, and signaling pathways. This review summarizes the formation mechanism, analysis, and optimization of structure and dynamics, as well as their functional applications in biomedicine and synthetic biology. Furthermore, the future development of sensors based on cpFP is envisioned to provide insights into the design of novel sensors.