Design strategies for fluorescent proteins/mimics and their applications in biosensing and bioimaging

Abstract 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.

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