Macromolecular Crowding Measurements with Genetically Encoded Probes Based on F örster Resonance Energy Transfer in Living Cells.

Macromolecular Crowding Measurements with Genetically Encoded Probes Based on Förster Resonance Energy Transfer in Living Cells. Methods Mol Biol. 2020;2175:169-180 Authors: Mouton SN, Veenhoff LM, Boersma AJ Abstract 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. PMID: 32681490 [PubMed - as supplied by publisher]
Source: Mol Biol Cell - Category: Molecular Biology Authors: Tags: Methods Mol Biol Source Type: research