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  Exchangeable HaloTag ligands for super-resolution fluorescence microscopy

Kompa, J., Bruins, J., Glogger, M., Wilhelm, J., Frei, M. S., Tarnawski, M., et al. (2022). Exchangeable HaloTag ligands for super-resolution fluorescence microscopy. Journal of the American Chemical Society, 145(5), 3075-3083. doi:10.1021/jacs.2c11969.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000C-83A0-A Version Permalink: https://hdl.handle.net/21.11116/0000-000C-8EF2-3
Genre: Journal Article

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 Creators:
Kompa, Julian1, Author           
Bruins, Jorick1, Author           
Glogger, Marius, Author
Wilhelm, Jonas1, Author           
Frei, Michelle S.1, Author           
Tarnawski, Miroslaw, Author
D´Este, Elisa2, Author           
Heilemann, Mike, Author
Hiblot, Julien1, Author           
Johnsson, Kai1, Author           
Affiliations:
1Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society, ou_2364732              
2Max Planck Institute for Medical Research, Max Planck Society, ou_1125545              

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 Abstract: The specific and covalent labeling of the protein HaloTag with fluorescent probes in living cells makes it a powerful tool for bioimaging. However, the irreversible attachment of the probe to HaloTag precludes imaging applications that require transient binding of the probe and comes with the risk of irreversible photobleaching. Here, we introduce exchangeable ligands for fluorescence labeling of HaloTag (xHTLs) that reversibly bind to HaloTag and that can be coupled to rhodamines of different colors. In stimulated emission depletion (STED) microscopy, probe exchange of xHTLs allows imaging with reduced photobleaching as compared to covalent HaloTag labeling. Transient binding of fluorogenic xHTLs to HaloTag fusion proteins enables points accumulation for imaging in nanoscale topography (PAINT) and MINFLUX microscopy. We furthermore introduce pairs of xHTLs and HaloTag mutants for dual-color PAINT and STED microscopy. xHTLs thus open up new possibilities in imaging across microscopy platforms for a widely used labeling approach.

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Language(s): eng - English
 Dates: 2022-11-102023-01-302022-02-08
 Publication Status: Issued
 Pages: 9
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1021/jacs.2c11969
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Title: Journal of the American Chemical Society
  Other : JACS
  Abbreviation : J. Am. Chem. Soc.
Source Genre: Journal
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Publ. Info: Washington, DC : American Chemical Society
Pages: - Volume / Issue: 145 (5) Sequence Number: - Start / End Page: 3075 - 3083 Identifier: ISSN: 0002-7863
CoNE: https://pure.mpg.de/cone/journals/resource/954925376870