Enhancing the photostability of red fluorescent proteins through FRET with Si-rhodamine for dynamic super-resolution fluorescence imaging.

ABSTRACT: Red fluorescent proteins (RFPs) are extensively utilized in biological imaging. However, their susceptibility to photobleaching restricts their effectiveness in super-resolution imaging where high photostability is crucial. In this study, we substantially improved the photostability of RFPs by incorporating a hybrid Förster resonance energy transfer (FRET) pair, utilizing RFPs as the energy donor and a photostable fluorophore, tetramethyl-Si-rhodamine (TMSiR), as the acceptor. TMSiR was selectively introduced through fusion with the HaloTag protein linked to the RFPs. We constructed a series of mApple/mCherry-TMSiR pairs with varying FRET efficiencies. Our findings reveal that higher FRET efficiency in the mApple/mCherry-TMSiR complexes correlates with enhanced photostability of RFPs. FRET

SUBMITTER: Zhou X 

PROVIDER: S-EPMC12067651 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature