PUBLICATION
Orthogonal fluorescent chemogenetic reporters for multicolor imaging
- Authors
- Tebo, A.G., Moeyaert, B., Thauvin, M., Carlon-Andres, I., Böken, D., Volovitch, M., Padilla-Parra, S., Dedecker, P., Vriz, S., Gautier, A.
- ID
- ZDB-PUB-200812-7
- 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.
Citation
Tebo, A.G., Moeyaert, B., Thauvin, M., Carlon-Andres, I., Böken, D., Volovitch, M., Padilla-Parra, S., Dedecker, P., Vriz, S., Gautier, A. (2020) Orthogonal fluorescent chemogenetic reporters for multicolor imaging. Nature Chemical Biology. 17(1):30-38.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping