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SNAP-tag2 for faster and brighter protein labeling

SNAP-tag is a powerful tool for labeling proteins with synthetic fluorophores in bioimaging. However, its utility in live-cell applications can be constrained by its relatively slow labeling kinetics and the limited cell permeability of its substrates. Here, we introduce improved labeling substrates...

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Hauptverfasser: Kühn, Stefanie (VerfasserIn) , Nasufović, Veselin (VerfasserIn) , Wilhelm, Jonas (VerfasserIn) , Kompa, Julian (VerfasserIn) , de Lange, Eline M. F. (VerfasserIn) , Lin, Yin-Hsi (VerfasserIn) , Egoldt, Cornelia (VerfasserIn) , Fischer, Jonas (VerfasserIn) , Lennoi, Artem (VerfasserIn) , Tarnawski, Miroslaw (VerfasserIn) , Reinstein, Jochen (VerfasserIn) , Vlijm, Rifka (VerfasserIn) , Hiblot, Julien (VerfasserIn) , Johnsson, Kai (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 03 July 2025
In: Nature chemical biology
Year: 2025, Jahrgang: 21, Heft: 11, Pages: 1754-1761
ISSN:1552-4469
DOI:10.1038/s41589-025-01942-z
Online-Zugang:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1038/s41589-025-01942-z
Verlag, lizenzpflichtig, Volltext: https://www.nature.com/articles/s41589-025-01942-z
Volltext
Verfasserangaben:Stefanie Kühn, Veselin Nasufovic, Jonas Wilhelm, Julian Kompa, Eline M. F. de Lange, Yin-Hsi Lin, Cornelia Egoldt, Jonas Fischer, Artem Lennoi, Miroslaw Tarnawski, Jochen Reinstein, Rifka Vlijm, Julien Hiblot, Kai Johnsson
Beschreibung
Zusammenfassung:SNAP-tag is a powerful tool for labeling proteins with synthetic fluorophores in bioimaging. However, its utility in live-cell applications can be constrained by its relatively slow labeling kinetics and the limited cell permeability of its substrates. Here, we introduce improved labeling substrates and an engineered SNAP-tag for faster labeling in vitro and in live cells. SNAP-tag2 presents a second-order rate constant with rhodamine substrates that approaches 107 s−1 M−1, a 100-fold improvement over the corresponding SNAP-tag-substrate pairs. When labeled with highly fluorogenic dyes, SNAP-tag2 also shows a fivefold increase in fluorescence brightness relative to currently used SNAP-tag. The increased labeling kinetics and brightness of SNAP-tag2 translate into greatly improved performance in various live-cell (super-resolution) imaging applications.
Beschreibung:Gesehen am 18.11.2025
Beschreibung:Online Resource
ISSN:1552-4469
DOI:10.1038/s41589-025-01942-z

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