期刊
ACS CHEMICAL BIOLOGY
卷 13, 期 9, 页码 2392-2397出版社
AMER CHEMICAL SOC
DOI: 10.1021/acschembio.8b00417
关键词
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资金
- European Research Council [ERC-2016-CoG-724705 FLUOSWITCH]
- Agence National de la Recherche [ANR-14-CE09-0002- 01]
- France BioImaging [ANR-10-INBS-04]
- Equipex Morphoscope 2 [ANR-11-EQPX-0029]
- program Investissements d'Avenir [ANR-10-IDEX-0001-02 PSL]
- Agence Nationale de la Recherche (ANR) [ANR-14-CE09-0002] Funding Source: Agence Nationale de la Recherche (ANR)
Fluorescent reporters are essential components for the design of optical biosensors that are able to image intracellular analytes in living cells. Herein, we describe the development of circularly permuted variants of Fluorescence-Activating and absorption-Shifting Tag (FAST) and demonstrate their potential as reporting module in biosensors. Circularly permutated FAST (cpFAST) variants allow one to condition the binding and activation of a fluorogenic ligand (and thus fluorescence) to analyte recognition by coupling them with analyte-binding domains. We demonstrated their use for biosensor design by generating multicolor plug-and-play fluorogenic biosensors for imaging the intracellular levels of Ca2+ in living mammalian cells in real time.
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