Halo-tag Labeling Technology and It’s Application in Plant Living Cell Imaging

doi:10.11983/CBB22227

Abstract

Abstract: The cell is the basic structural and functional unit of living organisms, its complex internal organizational structure, interaction and process dynamics determine the life form and life process of the entire organism. The study of cell structure and function in the living state is of great significance for exploring and grasping the essence of life. With the continuous advances in science and technology, a series of new live cell labeling technologies have been innovated. In recent years, the Halo-tag labeling technology has gradually been developed into a widely used new molecular labeling technology, playing an increasingly important role in the field of living cell imaging analysis and tracking research. It has been gradually applied in plant cell imaging. In this review, we focus on the definition, classification and development process of Halo-tag technology, and introduce the reaction mechanism of the dehydrohalogenation of Halo-tag labeling technology and the types of fluorescent ligands in detail. Finally, we introduce the latest progress of this technology in the application of living cell imaging in plants is emphatically expounded from three aspects: observing the fine localization of molecules, tracking the real-time motion of molecules and detecting the interactions between molecules. Taken together, this study provides theoretical foundation and technical support for the potential application of the subsequent Halo-tag labeling technology in plants.

Key words: new labeling technology, Halo-tag label, Halo-tag ligand, living cell imaging

Hongping Qian, Pengyun Luo, Shuai Liu, Changwen Xu, Jinhuan Yin, Yaning Cui, Jinxing Lin. Halo-tag Labeling Technology and It’s Application in Plant Living Cell Imaging[J]. Chinese Bulletin of Botany, 2023, 58(3): 475-485.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB22227

https://www.chinbullbotany.com/EN/Y2023/V58/I3/475

Figures/Tables 2

Figure 1 Halo-tag technology (A) Schematic showing Halo-tag labeling in living cells; (B) Chemical structural formulae of wild-type (WT) and mutant dehalogenases (green box indicates that aspartic acid (Asp) at position 106 of dehalogenase, which is the site where dehalogenation occurs and the intermediate ester is formed; red box indicates the difference between the wild-type dehalogenase and the mutant dehalogenase, where the wild-type dehalogenase is histidine (His) at position 272, while the mutant dehalogenase mutates to phenylalanine (Phe) at position 272); (C) Molecular model of the Halo-tag protein with a covalently bound TMR ligand (modified from England et al., 2015).

Table 1 Types of Halo-tag ligands for living cell imaging (modified from Benink and Urh, 2015)

配基类别	膜渗透性	激发/发射波长的峰值(nm)	是否需要清洗	化学结构式
TMR	细胞渗透性	555/585	是
diAcFAM	细胞渗透性	494/526	是
Oregon Green	细胞渗透性	496/516	是
Coumarin	细胞渗透性	353/434	是
Alexa Fluor? 488	非细胞渗透性	494/517	是
Alexa Fluor? 660	非细胞渗透性	663/690	是	结构图无法获得
TMRDirect?	细胞渗透性	555/585	否
R110Direct?	细胞渗透性	502/527	否

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