Dynamic multicolor protein labeling in living cells

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 to tune the fluorescence color of FAST from greenyellow to orange and red. Beyond allowing 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 selective detection of FAST-tagged proteins in cells expressing both green and red fluorescent species through two-color crosscorrelation, opening exciting prospects to overcome spectral crowding and push the frontiers of multiplexed imaging.



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Dynamic multicolor protein labeling in living cells
Chenge Li, Marie-Aude Plamont, Hanna L. Sladitschek, Vanessa Rodrigues, Isabelle Aujard, Pierre Neveu, Thomas Le Saux, Ludovic Jullien and Arnaud Gautier
Chem. Sci., 2017, Advance Article
DOI: 10.1039/C7SC01364G