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A far-red fluorescent chemogenetic reporter for in vivo molecular imaging

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Li, Chenge | Tebo, Alison, G | Thauvin, Marion | Plamont, Marie-Aude | Volovitch, Michel | Morin, Xavier | Vriz, Sophie | Gautier, Arnaud

Edité par CCSD ; Wiley-VCH Verlag -

International audience. Far-red emitting fluorescent labels are highly desirable for spectral multiplexing and deep tissue imaging. Here, we describe the generation of frFAST (far-red Fluorescence Activating and absorption Shifting Tag), a 14-kDa monomeric protein that forms a bright far-red fluorescent assembly with (4-hydroxy-3-methoxy-phenyl)allylidene rhodanine (HPAR-3OM). As HPAR-3OM is essentially nonfluorescent in solution and in cells, frFAST can be imaged with high contrast in presence of free HPAR-3OM, which allowed the rapid and efficient imaging of frFAST fusions in live cells, zebrafish embryo/larvae and chicken embryo. Beyond enabling genetic encoding of far-red fluorescence, frFAST allowed the design of a farred chemogenetic reporter of protein-protein interactions, demonstrating its great potential for the design of innovative far-red emitting biosensors.

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