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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.

SUBMITTER: Li C

PROVIDER: S-EPMC5618792 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature

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Dynamic multicolor protein labeling in living cells.

Li Chenge C Plamont Marie-Aude MA Sladitschek Hanna L HL Rodrigues Vanessa V Aujard Isabelle I Neveu Pierre P Le Saux Thomas T Jullien Ludovic L Gautier Arnaud A

Chemical science 20170530 8

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 ...[more]

PMID: 28970939

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