Recent advances in the design of small molecule-based FRET sensors for cell biology

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Fluorescence resonance energy transfer (FRET) has been used extensively as the designing principle for fluorescent sensor molecules. One of the most significant advantages of designing sensor molecules with FRET modulation is that it can enable ratiometric measurement in living cells, which reduces artifacts from microscopic imaging system. We describe the design strategy for the development of small-molecule FRET sensor in terms of avoiding close contact of donor fluorophore and acceptor fluorophore in aqueous solution. Furthermore, we introduce a strategy to design FRET sensors with a modulating overlap integral of donor and acceptor.