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Issue 8, 2017
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Dynamic multicolor protein labeling in living cells

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Abstract

Yellow Fluorescence-Activating and absorption-Shifting Tag (Y-FAST, hereafter called FAST) is a 14 kDa protein tag giving a bright green-yellow fluorescent complex upon interaction with the fluorogenic dye 4-hydroxy-3-methylbenzylidene rhodanine (HMBR). Here, we report a collection of fluorogens enabling tuning of the fluorescence color of FAST from green-yellow to orange and red. Beyond allowing the multicolor imaging of FAST-tagged proteins in live cells, these fluorogens enable dynamic color switching because of FAST’s reversible labeling. This unprecedented behavior allows for selective detection of FAST-tagged proteins in cells expressing both green and red fluorescent species through two-color cross-correlation, opening up exciting prospects to overcome spectral crowding and push the frontiers of multiplexed imaging.

Graphical abstract: Dynamic multicolor protein labeling in living cells

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Supplementary files

Article information


Submitted
27 Mar 2017
Accepted
20 May 2017
First published
30 May 2017

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2017,8, 5598-5605
Article type
Edge Article

Dynamic multicolor protein labeling in living cells

C. Li, M. Plamont, H. L. Sladitschek, V. Rodrigues, I. Aujard, P. Neveu, T. Le Saux, L. Jullien and A. Gautier, Chem. Sci., 2017, 8, 5598
DOI: 10.1039/C7SC01364G

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