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  Engineered HaloTag variants for fluorescence lifetime multiplexing

Frei, M. S., Tarnawski, M., Roberti, M. J., Koch, B., Hiblot, J., & Johnsson, K. (2022). Engineered HaloTag variants for fluorescence lifetime multiplexing. Nature Methods, 19, 65-70. doi:10.1038/s41592-021-01341-x.

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 Creators:
Frei, Michelle S.1, Author              
Tarnawski, Miroslaw2, Author              
Roberti, M. Julia, Author
Koch, Birgit1, Author              
Hiblot, Julien1, Author              
Johnsson, Kai1, Author              
Affiliations:
1Chemical Biology, Max Planck Institute for Medical Research, Max Planck Society, ou_2364732              
2Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Max Planck Society, ou_1497700              

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Free keywords: Chemical tools; Confocal microscopy; Fluorescence imaging; Fluorescent dyes; Proteins
 Abstract: Self-labeling protein tags such as HaloTag are powerful tools that can label fusion proteins with synthetic fluorophores for use in fluorescence microscopy. Here we introduce HaloTag variants with either increased or decreased brightness and fluorescence lifetime compared with HaloTag7 when labeled with rhodamines. Combining these HaloTag variants enabled live-cell fluorescence lifetime multiplexing of three cellular targets in one spectral channel using a single fluorophore and the generation of a fluorescence lifetime-based biosensor. Additionally, the brightest HaloTag variant showed up to 40% higher brightness in live-cell imaging applications.

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Language(s): eng - English
 Dates: 2021-03-312021-11-022021-12-162022-01
 Publication Status: Published in print
 Pages: 20
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1038/s41592-021-01341-x
 Degree: -

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Title: Nature Methods
  Other : Nature Methods
Source Genre: Journal
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Publ. Info: New York, NY : Nature Pub. Group
Pages: - Volume / Issue: 19 Sequence Number: - Start / End Page: 65 - 70 Identifier: ISSN: 1548-7091
CoNE: https://pure.mpg.de/cone/journals/resource/111088195279556