Fast and Luminous: CLIP-tag2
Johnsson, K.; Nasufovic, V.; Pispek, A.; Kuehn, S.; Wilhelm, J.; Bibrowski, M.; Fischer, J.; Koch, B.; Kompa, J.; Mao, R.; Tarnawski, M.; Hiblot, J.
Show abstract
CLIP-tag is a self-labeling protein tag used for the specific fluorescence labeling of proteins. However, its low labeling speed and the poor cell permeability of its substrates result in low labeling efficiencies in live-cell applications. Here, we introduce a substrate optimized for live-cell applications, as well as an engineered CLIP-tag variant, CLIP-tag2, which reacts with the new substrate almost 1000-fold faster than the original CLIP-tag-substrate pair. CLIP-tag2 fusion proteins can be specifically and efficiently fluorescently labeled in cells within minutes at nanomolar substrate concentrations, and can be multiplexed with other self-labeling tags such as SNAP-tag2 and HaloTag7. These advances establish CLIP-tag2 as a powerful tagging platform for high-performance live-cell bioimaging.
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