Dynamic multicolor protein labeling in living cells.
Chenge LiMarie-Aude PlamontHanna L SladitschekVanessa RodriguesIsabelle AujardPierre NeveuThomas Le SauxLudovic JullienArnaud GautierPublished in: Chemical science (2017)
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.
Keyphrases
- living cells
- single molecule
- induced apoptosis
- fluorescent probe
- cell cycle arrest
- high resolution
- signaling pathway
- quantum dots
- label free
- amino acid
- endoplasmic reticulum stress
- flow cytometry
- cell death
- protein protein
- optical coherence tomography
- energy transfer
- single cell
- oxidative stress
- mass spectrometry
- magnetic resonance
- photodynamic therapy
- aqueous solution
- sensitive detection