Engineered HaloTag variants for fluorescence lifetime multiplexing

Michelle S. Frei

doi:10.1038/s41592-021-01341-x

2021-12-16

DOI：10.1038/s41592-021-01341-x

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摘要

En 中文

HaloTag variants offer distinct brightness and fluorescence lifetimes compared with HaloTag7 when labeled with rhodamines. These variants were used for multiplexed imaging with a single fluorophore and to create lifetime-based cell cycle indicators. 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.

Keyword:

SMALL MOLECULES

GENERAL-METHOD

PROTEIN

DYNAMICS

PROBES

DYES

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期刊

Nature Methods

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32.1

论文数:

7.2K

被引数:

12.7W

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Ecole Polytechnique Federale de Lausanne

学者数:

1.7W

论文数: 1.3W

被引数: 25

Max Planck Society

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8.2W

论文数: 7.7W

被引数: 3.3W

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