ZFIN ID: ZDB-PUB-170214-57
Small fluorescence-activating and absorption-shifting tag for tunable protein imaging in vivo
Plamont, M.A., Billon-Denis, E., Maurin, S., Gauron, C., Pimenta, F.M., Specht, C.G., Shi, J., Quérard, J., Pan, B., Rossignol, J., Moncoq, K., Morellet, N., Volovitch, M., Lescop, E., Chen, Y., Triller, A., Vriz, S., Le Saux, T., Jullien, L., Gautier, A.
Date: 2016
Source: Proceedings of the National Academy of Sciences of the United States of America   113: 497-502 (Journal)
Registered Authors: Gauron, Carole, Triller, Antoine, Vriz, Sophie
Keywords: directed evolution, fluorescence imaging, fluorogenic ligand
MeSH Terms:
  • Absorption, Radiation*
  • Animals
  • Cell Survival
  • Embryo, Nonmammalian/metabolism
  • Fluorescence
  • Genetic Engineering
  • HeLa Cells
  • Humans
  • Imaging, Three-Dimensional*
  • Recombinant Fusion Proteins/metabolism*
  • Staining and Labeling
  • Subcellular Fractions/metabolism
  • Zebrafish/embryology
PubMed: 26711992 Full text @ Proc. Natl. Acad. Sci. USA
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ABSTRACT
This paper presents Yellow Fluorescence-Activating and absorption-Shifting Tag (Y-FAST), a small monomeric protein tag, half as large as the green fluorescent protein, enabling fluorescent labeling of proteins in a reversible and specific manner through the reversible binding and activation of a cell-permeant and nontoxic fluorogenic ligand (a so-called fluorogen). A unique fluorogen activation mechanism based on two spectroscopic changes, increase of fluorescence quantum yield and absorption red shift, provides high labeling selectivity. Y-FAST was engineered from the 14-kDa photoactive yellow protein by directed evolution using yeast display and fluorescence-activated cell sorting. Y-FAST is as bright as common fluorescent proteins, exhibits good photostability, and allows the efficient labeling of proteins in various organelles and hosts. Upon fluorogen binding, fluorescence appears instantaneously, allowing monitoring of rapid processes in near real time. Y-FAST distinguishes itself from other tagging systems because the fluorogen binding is highly dynamic and fully reversible, which enables rapid labeling and unlabeling of proteins by addition and withdrawal of the fluorogen, opening new exciting prospects for the development of multiplexing imaging protocols based on sequential labeling.
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