A Far-Red Emitting 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

doi:10.1002/anie.202006576

A Far-Red Emitting Fluorescent Chemogenetic Reporter for In Vivo Molecular Imaging

Angew Chem Int Ed Engl. 2020 Oct 5;59(41):17917-17923. doi: 10.1002/anie.202006576. Epub 2020 Aug 18.

Authors

Chenge Li^{1

2

3}, Alison G Tebo^{1

4}, Marion Thauvin^{5

6}, Marie-Aude Plamont¹, Michel Volovitch^{5

7}, Xavier Morin⁸, Sophie Vriz^{5

9}, Arnaud Gautier^{1

4

10}

Affiliations

¹ PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS, 75005, Paris, France.
² Present address: Department of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, China.
³ State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, China.
⁴ Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des biomolécules, LBM, 75005, Paris, France.
⁵ Center for Interdisciplinary Research in Biology (CIRB), Collège de France, CNRS, INSERM, Université PSL, Paris, France.
⁶ Sorbonne Université, Paris, France.
⁷ École Normale Supérieure, Université PSL, Department of biology, Paris, France.
⁸ Institut de Biologie de l'ENS (IBENS), École Normale Supérieure, CNRS, INSERM, Université PSL, 75005, Paris, France.
⁹ Université de Paris, Faculty of Science, 75006, Paris, France.
¹⁰ Institut Universitaire de France, France.

PMID: 32568417
DOI: 10.1002/anie.202006576

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 embryos. Beyond enabling the 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.

Keywords: biosensors; chemogenetic reporters; chromophores; far-red fluorescence; in vivo imaging.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Chick Embryo
Fluorescent Dyes / chemistry*
HeLa Cells
Humans
Molecular Imaging / methods*
Protein Binding
Zebrafish / embryology

Substances

Fluorescent Dyes