Saimi Dilizhatai, Chen Zhixing
College of Future Technology Institute of Molecular Medicine National Biomedical Imaging Center Beijing Key Laboratory of Cardiometabolic Molecular Medicine Peking University Beijing China.
Peking-Tsinghua Center for Life Science Academy for Advanced Interdisciplinary Studies Peking University Beijing China.
Smart Mol. 2023 Aug 28;1(2):e20230002. doi: 10.1002/smo.20230002. eCollection 2023 Sep.
Imaging proteins with high resolution is crucial for studying cellular physiology and pathology. Fluorescence imaging is a privileged method to visualize proteins with subcellular precision in live cells. In recent years, there has been a tremendous advance in the field of fluorescent dyes that are optically more sophisticated than genetically-encodable fluorescent proteins. In this review, we aim to discuss modern bioconjugation methods to specifically incorporate these dyes into protein-of-interests. We focus on advances in live-cell labeling strategies and fluorescent probes, especially the HaloTag, SNAP-tag, TMP-tag, and unnatural amino acid systems and their applications. These protein labeling methods, along with cutting-edge dyes and novel microscopy methods, have become the infrastructure for biological research in the era of super-resolution imaging.
以高分辨率成像蛋白质对于研究细胞生理学和病理学至关重要。荧光成像是一种在活细胞中以亚细胞精度可视化蛋白质的特权方法。近年来，在荧光染料领域取得了巨大进展，这些染料在光学上比基因编码的荧光蛋白更为复杂。在本综述中，我们旨在讨论将这些染料特异性掺入目标蛋白质的现代生物共轭方法。我们重点关注活细胞标记策略和荧光探针的进展，特别是卤代标签、SNAP标签、TMP标签和非天然氨基酸系统及其应用。这些蛋白质标记方法，连同前沿染料和新型显微镜方法，已成为超分辨率成像时代生物学研究的基础设施。
