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

Chenge Li, Alison G. Tebo, Marion Thauvin, Marie-Aude Plamont, Michel Volovitch, Xavier Morin, Sophie Vriz, View ORCID ProfileArnaud Gautier
doi: https://doi.org/10.1101/2020.04.04.022145
Chenge Li
1PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS, 75005 Paris, France
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Alison G. Tebo
1PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS, 75005 Paris, France
2Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des biomolécules, LBM, 75005 Paris, France
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Marion Thauvin
3Center for Interdisciplinary Research in Biology (CIRB), Collège de France, CNRS, INSERM, Université PSL, Paris, France
4Sorbonne Université, Paris, France
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Marie-Aude Plamont
1PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS, 75005 Paris, France
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Michel Volovitch
3Center for Interdisciplinary Research in Biology (CIRB), Collège de France, CNRS, INSERM, Université PSL, Paris, France
5École Normale Supérieure, Université PSL, Department of biology, Paris, France
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Xavier Morin
6Institut de Biologie de l’ENS (IBENS), École Normale Supérieure, CNRS, INSERM, Université PSL, 75005 Paris, France
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Sophie Vriz
3Center for Interdisciplinary Research in Biology (CIRB), Collège de France, CNRS, INSERM, Université PSL, Paris, France
7Université de Paris, Faculty of Science - 75006 Paris, France
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Arnaud Gautier
1PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS, 75005 Paris, France
2Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des biomolécules, LBM, 75005 Paris, France
8Institut Universitaire de France
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  • ORCID record for Arnaud Gautier
  • For correspondence: arnaud.gautier@sorbonne-universite.fr
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Abstract

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 non-fluorescent 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 far-red chemogenetic reporter of protein-protein interactions, demonstrating its great potential for the design of innovative far-red emitting biosensors.

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Posted April 05, 2020.
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A far-red fluorescent chemogenetic reporter for in vivo molecular imaging
Chenge Li, Alison G. Tebo, Marion Thauvin, Marie-Aude Plamont, Michel Volovitch, Xavier Morin, Sophie Vriz, Arnaud Gautier
bioRxiv 2020.04.04.022145; doi: https://doi.org/10.1101/2020.04.04.022145
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A far-red fluorescent chemogenetic reporter for in vivo molecular imaging
Chenge Li, Alison G. Tebo, Marion Thauvin, Marie-Aude Plamont, Michel Volovitch, Xavier Morin, Sophie Vriz, Arnaud Gautier
bioRxiv 2020.04.04.022145; doi: https://doi.org/10.1101/2020.04.04.022145

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