An expanded palette of fluorogenic HaloTag probes with enhanced contrast for targeted cellular imaging

Archive ouverte

Bachollet, Sylvestre | Shpinov, Yuriy | Broch, Fanny | Benaissa, Hela | Gautier, Arnaud | Pietrancosta, Nicolas | Mallet, Jean-Maurice | Dumat, Blaise

Edité par CCSD ; Royal Society of Chemistry -

International audience. We report the development of HaloTag fluorogens based on dipolar flexible molecular rotor structures. By modulating the electron donating and withdrawing groups, we have tuned the absorption and emission wavelengths to design a palette of fluorogens with emissions spanning the green to red range, opening new possibilities for multicolor imaging. The probes were studied in glycerol and in presence of HaloTag and exhibited good fluorogenic properties thanks to a viscosity-sensitive emission. In live-cell confocal imaging, the fluorogens yielded only a very low non-specific signal that enabled wash-free targeted imaging of intracellular organelles and proteins with good contrast. Combining experimental studies and theoretical investigation of the protein/fluorogen complexes by molecular dynamics, these results offer new insight into the design of molecular rotor-based fluorogenic HaloTag probes in order to improve reaction rates and the imaging contrast.

Suggestions

Du même auteur

Fluorogenic and genetic targeting of a red-emitting molecular calcium indicator

Archive ouverte | Bachollet, Sylvestre | CCSD

International audience. The fluorogenic activation of a calcium molecular probe via reaction with HaloTag yields a fluorescent AND logic gate enabling selective and wash-free calcium imaging in genetically-defined o...

Genetic Targeting of Solvatochromic Dyes for Probing Nanoscale Environments of Proteins in Organelles

Archive ouverte | Pelletier, Rémi | CCSD

International audience. A variety of protein tags are available for genetically encoded protein labelling, which allow their precise localization and tracking inside the cells. A new dimension in protein imaging can...

Fluorogenic Protein Probes with Red and Near‐Infrared Emission for Genetically Targeted Imaging

Archive ouverte | Bachollet, Sylvestre | CCSD

International audience. Fluorogenic probes are important tools to image proteins with high contrast and no wash protocols. In this work, we rationally designed and synthesized a small set of four protein fluorogens ...

Chargement des enrichissements...