ZFIN ID: ZDB-PUB-200812-7
Orthogonal fluorescent chemogenetic reporters for multicolor imaging
Tebo, A.G., Moeyaert, B., Thauvin, M., Carlon-Andres, I., Böken, D., Volovitch, M., Padilla-Parra, S., Dedecker, P., Vriz, S., Gautier, A.
Date: 2020
Source: Nature Chemical Biology   17(1): 30-38 (Journal)
Registered Authors: Gautier, Aude, Vriz, Sophie
Keywords: none
MeSH Terms:
  • Animals
  • Benzylidene Compounds/chemistry
  • Biosensing Techniques*
  • COS Cells
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Color
  • Escherichia coli/genetics
  • Escherichia coli/metabolism
  • Fluorescent Dyes/chemistry*
  • Fluorescent Dyes/metabolism
  • Gene Expression
  • Molecular Biology/methods*
  • Oligonucleotides/genetics
  • Oligonucleotides/metabolism
  • Optical Imaging/methods*
  • Plasmids/chemistry*
  • Plasmids/metabolism
  • Protein Engineering
  • Recombinant Proteins/chemistry
  • Recombinant Proteins/genetics
  • Recombinant Proteins/metabolism
  • Saccharomyces cerevisiae/genetics
  • Saccharomyces cerevisiae/metabolism
  • Staining and Labeling/methods*
  • Zebrafish
PubMed: 32778846 Full text @ Nat. Chem. Biol.
Spectrally separated fluorophores allow the observation of multiple targets simultaneously inside living cells, leading to a deeper understanding of the molecular interplay that regulates cell function and fate. Chemogenetic systems combining a tag and a synthetic fluorophore provide certain advantages over fluorescent proteins since there is no requirement for chromophore maturation. Here, we present the engineering of a set of spectrally orthogonal fluorogen-activating tags based on the fluorescence-activating and absorption shifting tag (FAST) that are compatible with two-color, live-cell imaging. The resulting tags, greenFAST and redFAST, demonstrate orthogonality not only in their fluorogen recognition capabilities, but also in their one- and two-photon absorption profiles. This pair of orthogonal tags allowed the creation of a two-color cell cycle sensor capable of detecting very short, early cell cycles in zebrafish development and the development of split complementation systems capable of detecting multiple protein-protein interactions by live-cell fluorescence microscopy.