Engineered HaloTag variants for fluorescence lifetime multiplexing

Frei, Michelle S.; Tarnawski, Miroslaw; Roberti, M. Julia; Koch, Birgit; Hiblot, Julien; Johnsson, Kai

doi:10.1038/s41592-021-01341-x

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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: Submitted: 2021-03-31Accepted: 2021-11-02Published Online: 2021-12-16Date issued: 2022-01

Publication Status: Issued

Pages: 20

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Table of Contents: -

Rev. Type: Peer

Identifiers: DOI: 10.1038/s41592-021-01341-x

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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