A high-affinity split-HaloTag for live-cell protein labeling

doi:10.1038/s41467-026-71032-8

2026-03-25

DOI：10.1038/s41467-026-71032-8

原文链接

查看原文

摘要

En 中文

我们介绍了一种高亲和力分裂型HaloTag，由一个短肽标签（Hpep，14个残基）和一个大的无活性片段（cpHaloΔ3）组成。Hpep与cpHaloΔ3自发结合，亲和力为纳摩尔级别，能够实现后续用荧光HaloTag配体进行标记。Hpep的小尺寸促进了无需克隆的内源蛋白标记，使用CRISPR/Cas9技术，Hpep标记蛋白的互补可以在活细胞中通过共表达cpHaloΔ3实现，在固定细胞中通过孵育cpHaloΔ3实现。该方法与扩展显微镜和活细胞STED成像等先进显微技术兼容。此外，Hpep的变体能够调节标记荧光团的谱学特性，通过荧光寿命显微成像技术实现两种不同Hpep标记蛋白的同时成像。总之，我们高亲和力的分裂型HaloTag是活细胞成像和化学生物学多样应用中的一个强大且多功能的工具。Lin及其同事介绍了用于蛋白标记和多标记的高亲和力分裂型HaloTag对。该多功能系统允许使用包括活细胞超分辨率显微成像在内的多种成像模式进行蛋白可视化。

Keyword:

Fluorescence imaging

Protein design

Science

Humanities and Social Sciences

multidisciplinary

AI总结

对已上传原文的论文进行重点信息的提取，主要内容包括：简要概述、研究摘要、背景介绍、关键亮点、图文解析、展望与总结。

期刊

Nature Communications

IF:

15.7

论文数:

9.4W

被引数:

91.2W

机构

Max Planck Institute for Medical Research

学者数:

论文数: 24

被引数: 1.5K

引用论文

IF0

PREAI

Overview of Affinity Tags for Protein Purification

Current Protocols in Protein Science

IF0

2018-02-16

OAAI

Michelle E. Kimple; Allison L. Brill; Renee L. Pasker

A modular chemigenetic calcium indicator for multiplexed in vivo functional imaging

NATURE METHODS

IF32.1

2024-09-20

OAAI

Farrants, Helen; Shuai, Yichun; Lemon, William C.; Monroy Hernandez, Christian; Zhang, Deng; Yang, Shang; Patel, Ronak; Qiao, Guanda; Frei, Michelle S.; Plutkis, Sarah E.; Grimm, Jonathan B.; Hanson, Timothy L.; Tomaska, Filip; Turner, Glenn C.; Stringer, Carsen; Keller, Philipp J.; Beyene, Abraham G.; Chen, Yao; Liang, Yajie; Lavis, Luke D.; Schreiter, Eric R.

Rapid and Reversible Knockdown of Endogenously Tagged Endosomal Proteins via an Optimized HaloPROTAC Degrader

ACS CHEMICAL BIOLOGY

IF3.8

2019-04-12

OAAI

Tovell, Hannah; Testa, Andrea; Maniaci, Chiara; Zhou, Houjiang; Prescott, Alan R.; Macartney, Thomas; Ciulli, Alessio; Alessi, Dario R.

Imaging cellular ultrastructures using expansion microscopy (U-ExM)

NATURE METHODS

IF32.1

2018-12-17

271

OAAI

Gambarotto, Davide; Zwettler, Fabian U.; Le Guennec, Maeva; Schmidt-Cernohorska, Marketa; Fortun, Denis; Borgers, Susanne; Heine, Joern; Schloetel, Jan-Gero; Reuss, Matthias; Unser, Michael; Boyden, Edward S.; Sauer, Markus; Hamel, Virginie; Guichard, Paul

Specific Covalent Labeling of Recombinant Protein Molecules Inside Live Cells

Science

IF0

1998-07-10

PREAI

B. Albert Griffin; Stephen R. Adams; Roger Y. Tsien

A gp41 MPER-specific Llama VHH Requires a Hydrophobic CDR3 for Neutralization but not for Antigen Recognition

PLOS PATHOGENS

IF4.9

2013-03-07

OAAI

Hulsik, David Lutje; Liu, Ying-ying; Strokappe, Nika M.; Battella, Simone; El Khattabi, Mohamed; McCoy, Laura E.; Sabin, Charles; Hinz, Andreas; Hock, Miriam; Macheboeuf, Pauline; Bonvin, Alexandre M. J. J.; Langedijk, Johannes P. M.; Davis, David; Quigley, Anna Forsman; Aasa-Chapman, Marlen M. I.; Seaman, Michael S.; Ramos, Alejandra; Poignard, Pascal; Favier, Adrien; Simorre, Jean-Pierre; Weiss, Robin A.; Verrips, C. Theo; Weissenhorn, Winfried; Rutten, Lucy

A new method for post-translationally labeling proteins in live cells for fluorescence imaging and tracking

Protein Engineering, Design and Selection

IF0

2017-12-06

OAAI

M Hinrichsen; M Lenz; J M Edwards; O K Miller; S G J Mochrie; P S Swain; U Schwarz-Linek; L Regan

Split-HaloTag imaging assay for sophisticated microscopy of protein-protein interactions in planta

PLANT COMMUNICATIONS

IF11.6

2021-09-01

OAAI

Minner-Meinen, Rieke; Weber, Jan-Niklas; Albrecht, Andreas; Matis, Rainer; Behnecke, Maria; Tietge, Cindy; Frank, Stefan; Schulze, Jutta; Buschmann, Henrik; Walla, Peter Jomo; Mendel, Ralf-R.; Haensch, Robert; Kaufholdt, David

Multi-Color Single-Molecule Imaging Uncovers Extensive Heterogeneity in mRNA Decoding 多色单分子成像揭示了mRNA解码中的广泛异质性

Cell

IF0

2019-07-01

OAAI

Sanne Boersma; Deepak Khuperkar; Bram M.P. Verhagen; Stijn Sonneveld; Jonathan B. Grimm; Luke D. Lavis; Marvin E. Tanenbaum

查看更多内容