Porzberg Nicola, Gries Klara, Johnsson Kai
Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany; email:
Institute of Chemical Sciences and Engineering (ISIC), École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
Annu Rev Biochem. 2025 Jun;94(1):29-58. doi: 10.1146/annurev-biochem-030222-121016. Epub 2025 Mar 19.
The visualization and manipulation of proteins in live cells are critical for studying complex biological processes. Self-labeling proteins do so by enabling the specific and covalent attachment of synthetic probes, offering unprecedented flexibility in the chemical labeling of proteins in live cells and in vivo. By combining the excellent photophysical properties of synthetic dyes with genetic targetability, these tags provide a modular and innovative toolbox for live-cell and high-resolution fluorescence imaging. In this review, we explore the development and diverse applications of the key self-labeling protein technologies, HaloTag7, SNAP-tag, and CLIP-tag, as well as the covalent trimethoprim (TMP)-tag. We discuss recent innovations in both protein engineering and substrate design that have introduced new functionalities to enable multiplexed imaging, super-resolution microscopy, and the design of novel biosensors and recorders.
活细胞中蛋白质的可视化和操控对于研究复杂的生物过程至关重要。自标记蛋白通过实现合成探针的特异性共价连接来做到这一点，为活细胞和体内蛋白质的化学标记提供了前所未有的灵活性。通过将合成染料出色的光物理特性与基因靶向性相结合，这些标签为活细胞和高分辨率荧光成像提供了一个模块化且创新的工具箱。在本综述中，我们探讨了关键的自标记蛋白技术——HaloTag7、SNAP标签、CLIP标签以及共价三甲氧苄氨嘧啶（TMP）标签的发展和多样应用。我们讨论了蛋白质工程和底物设计方面的最新创新，这些创新引入了新功能，以实现多重成像、超分辨率显微镜以及新型生物传感器和记录器的设计。
