Fluorescent proteins (FPs) are powerful and fundamental tools in bio-analytical research, ranging from tracking biological molecules to monitoring dynamic processes in living cells. Currently, great efforts have been made on structurally engineering of FPs with new fluorescent signal-switch mechanisms via rational or semi-rational design. In this review, we summarize a variety of strategies for redesigning FPs, including circularly permuted FPs, split FPs, supercharged FPs, and unnatural amino acid-containing FPs, as well as RNA/DNA mimics of FPs. We focus on the design strategies and target-responsive mechanisms of the engineered FPs and their current applications in biosensing and bioimaging. The recent advances summarized here reveal that the great flexibility of the structure and the outstanding designability endow FPs with remarkable potential as versatile sensing elements for biochemical analysis.

荧光蛋白（FPs）是生物分析研究中强大而基础的工具，范围从跟踪生物分子到监测活细胞中的动态过程。当前，已经在通过合理或半合理设计的具有新的荧光信号切换机制的FP的结构工程上做出了巨大的努力。在这篇综述中，我们总结了用于重新设计FP的多种策略，包括圆形排列的FP，分裂的FP，增压的FP和非天然氨基酸的FP，以及FP的RNA / DNA模拟物。我们关注工程化FP的设计策略和目标响应机制及其在生物传感和生物成像中的当前应用。