Design of Bright Chemogenetic Reporters Based on the Combined Engineering of Fluorogenic Molecular Rotors and of the HaloTag Protein - Normal and Pathologic Glutamatergic Neurons Access content directly
Journal Articles Chemistry - A European Journal Year : 2024

Design of Bright Chemogenetic Reporters Based on the Combined Engineering of Fluorogenic Molecular Rotors and of the HaloTag Protein

Abstract

The combination of fluorogenic probes (fluorogens) and self‐labeling protein tags represent a promising tool for imaging biological processes with high specificity but it requires the adequation between the fluorogen and its target to ensure a good activation of its fluorescence. In this work, we report a strategy to develop molecular rotors that specifically target HaloTag with a strong enhancement of their fluorescence. The divergent design facilitates the diversification of the structures to tune the photophysical and cellular properties. Four bright fluorogens with emissions ranging from green to red were identified and applied in wash‐free live cell imaging experiments with good contrast and selectivity. A HaloTag mutant adapted from previous literature reports was also tested and shown to further improve the brightness and reaction rate of the most promising fluorogen of the series both in vitro and in cells. This work opens new possibilities to develop bright chemogenetic reporters with diverse photophysical and biological properties by exploring a potentially large chemical space of simple dipolar fluorophores in combination with protein engineering.
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Origin Publication funded by an institution

Dates and versions

hal-04569035 , version 1 (06-05-2024)

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Justine Coïs, Sylvestre Bachollet, Louis Sanchez, Nicolas Pietrancosta, Vincent Vialou, et al.. Design of Bright Chemogenetic Reporters Based on the Combined Engineering of Fluorogenic Molecular Rotors and of the HaloTag Protein. Chemistry - A European Journal, inPress, ⟨10.1002/chem.202400641⟩. ⟨hal-04569035⟩
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