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: | , , , , , , , , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
03 July 2025
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| 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 
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| 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 |
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| 520 | |a 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. | ||
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