Design strategies and applications of fluorescent protein-based probes.

The discovery of fluorescent proteins has revolutionized cell biology research. By fusing fluorescent proteins with target proteins, fluorescent biosensors can be constructed to enable real-time monitoring of dynamic cellular events in live cells and organisms. The unique physicochemical properties of fluorescent proteins, such as spectral range, chromophore maturation speed, pH sensitivity, and stability, offer diverse options for probe design. Researchers have developed various fluorescent biosensors based on these properties to monitor distinct molecular events. This review systematically summarizes the main strategies for designing fluorescent protein probes and highlights their typical applications in biological research. It provides a reference for developing more efficient and specialized fluorescent probes to address complex biological questions.