Genetically encoded Förster resonance energy transfer (FRET)-based probes allow a sensitive readout for different or specific parameters in the living cell. We previously demonstrated how FRET-based probes could quantify macromolecular crowding with high spatio-temporal resolution and under various conditions. Here, we present a protocol developed for the use of FRET-based crowding probes in baker’s yeast, but the general considerations also apply to other species, as well as other FRET-based sensors. This method allows straightforward detection of macromolecular crowding under challenging conditions often presented by living cells.
CITATION STYLE
Mouton, S. N., Veenhoff, L. M., & Boersma, A. J. (2020). Macromolecular crowding measurements with genetically encoded probes based on förster resonance energy transfer in living cells. In Methods in Molecular Biology (Vol. 2175, pp. 169–180). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0763-3_12
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