Engineered HaloTag variants for fluorescence lifetime multiplexing

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...

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Autores principales: Frei, Michelle S., Tarnawski, Miroslaw, Roberti, M. Julia, Koch, Birgit, Hiblot, Julien, Johnsson, Kai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group US 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8748199/
https://www.ncbi.nlm.nih.gov/pubmed/34916672
http://dx.doi.org/10.1038/s41592-021-01341-x
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author Frei, Michelle S.
Tarnawski, Miroslaw
Roberti, M. Julia
Koch, Birgit
Hiblot, Julien
Johnsson, Kai
author_facet Frei, Michelle S.
Tarnawski, Miroslaw
Roberti, M. Julia
Koch, Birgit
Hiblot, Julien
Johnsson, Kai
author_sort Frei, Michelle S.
collection PubMed
description 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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spelling pubmed-87481992022-01-20 Engineered HaloTag variants for fluorescence lifetime multiplexing Frei, Michelle S. Tarnawski, Miroslaw Roberti, M. Julia Koch, Birgit Hiblot, Julien Johnsson, Kai Nat Methods Article 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. Nature Publishing Group US 2021-12-16 2022 /pmc/articles/PMC8748199/ /pubmed/34916672 http://dx.doi.org/10.1038/s41592-021-01341-x Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Frei, Michelle S.
Tarnawski, Miroslaw
Roberti, M. Julia
Koch, Birgit
Hiblot, Julien
Johnsson, Kai
Engineered HaloTag variants for fluorescence lifetime multiplexing
title Engineered HaloTag variants for fluorescence lifetime multiplexing
title_full Engineered HaloTag variants for fluorescence lifetime multiplexing
title_fullStr Engineered HaloTag variants for fluorescence lifetime multiplexing
title_full_unstemmed Engineered HaloTag variants for fluorescence lifetime multiplexing
title_short Engineered HaloTag variants for fluorescence lifetime multiplexing
title_sort engineered halotag variants for fluorescence lifetime multiplexing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8748199/
https://www.ncbi.nlm.nih.gov/pubmed/34916672
http://dx.doi.org/10.1038/s41592-021-01341-x
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