Cover Image

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...

Full description

Saved in:
Bibliographic Details
Main Authors: Kühn, Stefanie (Author) , Nasufović, Veselin (Author) , Wilhelm, Jonas (Author) , Kompa, Julian (Author) , de Lange, Eline M. F. (Author) , Lin, Yin-Hsi (Author) , Egoldt, Cornelia (Author) , Fischer, Jonas (Author) , Lennoi, Artem (Author) , Tarnawski, Miroslaw (Author) , Reinstein, Jochen (Author) , Vlijm, Rifka (Author) , Hiblot, Julien (Author) , Johnsson, Kai (Author)
Format: Article (Journal)
Language:English
Published: 03 July 2025
In: Nature chemical biology
Year: 2025, Volume: 21, Issue: 11, Pages: 1754-1761
ISSN:1552-4469
DOI:10.1038/s41589-025-01942-z
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1038/s41589-025-01942-z
Verlag, lizenzpflichtig, Volltext: https://www.nature.com/articles/s41589-025-01942-z
Get full text
Author Notes: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
Description
Summary: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.
Item Description:Gesehen am 18.11.2025
Physical Description:Online Resource
ISSN:1552-4469
DOI:10.1038/s41589-025-01942-z

Similar Items