Issue 8, 2017

Dynamic multicolor protein labeling in living cells

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

Supplementary files

Article information

Article type
Edge Article
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
Creative Commons BY license

Chem. Sci., 2017,8, 5598-5605

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

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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