期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 23, 期 8, 页码 -出版社
MDPI
DOI: 10.3390/ijms23084396
关键词
FRET; fluorogen-activating proteins; FAPs
资金
- Ministry of Science and Higher Education of the Russian Federation [075-15-2020-773]
This study introduces a modulated FRET acceptor that simplifies one-channel FRET measurement by using a cell-permeable chromophore to decrease the fluorescence of the donor-fluorescent protein. The chemically modulated FRET acceptor is compatible with live-cell experiments and allows for prolonged time-lapse experiments with dynamic energy transfer evaluation.
A substantial number of genetically encoded fluorescent sensors rely on the changes in FRET efficiency between fluorescent cores, measured in ratiometric mode, with acceptor photobleaching or by changes in fluorescence lifetime. We report on a modulated FRET acceptor allowing for simplified one-channel FRET measurement based on a previously reported fluorogen-activating protein, DiB1. Upon the addition of the cell-permeable chromophore, the fluorescence of the donor-fluorescent protein mNeonGreen decreases, allowing for a simplified one-channel FRET measurement. The reported chemically modulated FRET acceptor is compatible with live-cell experiments and allows for prolonged time-lapse experiments with dynamic energy transfer evaluation.
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