Engineering of a fluorescent chemogenetic reporter with tunable color for advanced live-cell imaging

• Published in: Nature communications Vol. 12; no. 1; p. 6989
• Main Authors: Benaissa, Hela, Ounoughi, Karim, Aujard, Isabelle, Fischer, Evelyne, Goïame, Rosette, Nguyen, Julie, Tebo, Alison G., Li, Chenge, Le Saux, Thomas, Bertolin, Giulia, Tramier, Marc, Danglot, Lydia, Pietrancosta, Nicolas, Morin, Xavier, Jullien, Ludovic, Gautier, Arnaud
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30-11-2021; Nature Publishing Group; Nature Portfolio
• Subjects: 14; 14/19; 14/33; 14/34; 14/35; 14/63; 631/1647/527/2047; 631/80/2373/2238; 631/92/96; Animals; Biocompatibility; Biocompatible Materials; Bioengineering; Biosensing Techniques; Biosensors; Chemical Sciences; Chromophores; Color; Coloring Agents; Diagnostic Imaging - methods; Directed evolution; Electronics; Energy transfer; Engineering; Female; Fluorescence; Fluorescence Resonance Energy Transfer; Fluorescent Dyes; Green Fluorescent Proteins; Humanities and Social Sciences; Image resolution; Imaging; Imaging techniques; Life Sciences; Male; multidisciplinary; Mutation; Neurons; Neurosciences; Optical properties; or physical chemistry; Organic chemistry; Organisms; Physiology; Protein Engineering - methods; Proteins; Rats; Rats, Sprague-Dawley; Science; Science (multidisciplinary); Spectra; Theoretical and; Visible spectrum
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-27334-0

Biocompatible fluorescent reporters with spectral properties spanning the entire visible spectrum are indispensable tools for imaging the biochemistry of living cells and organisms in real time. Here, we report the engineering of a fluorescent chemogenetic reporter with tunable optical and spectral properties. A collection of fluorogenic chromophores with various electronic properties enables to generate bimolecular fluorescent assemblies that cover the visible spectrum from blue to red using a single protein tag engineered and optimized by directed evolution and rational design. The ability to tune the fluorescence color and properties through simple molecular modulation provides a broad experimental versatility for imaging proteins in live cells, including neurons, and in multicellular organisms, and opens avenues for optimizing Förster resonance energy transfer (FRET) biosensors in live cells. The ability to tune the spectral properties and fluorescence performance enables furthermore to match the specifications and requirements of advanced super-resolution imaging techniques.
Fluorescent reporters spanning the visible spectrum are needed for imaging live cells and organisms. Here the authors report a collection of fluorogenic chromophores that cover the visible spectrum from blue to red using a single engineered and optimised protein tag.