|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.
|Source:||Nature Chemical Biology 17(1): 30-38 (Journal)|
|Registered Authors:||Gautier, Aude, Vriz, Sophie|
|PubMed:||32778846 Full text @ Nat. Chem. Biol.|
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.
ABSTRACTSpectrally 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.
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