Beyond the green fluorescent protein: biomolecular reporters for anaerobic and deep-tissue imaging.
Beyond the green fluorescent protein: biomolecular reporters for anaerobic and deep-tissue imaging.
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DOI：
10.1021/acs.bioconjchem.9b00688
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发表时间：
2019-12
影响因子：
4.7
通讯作者：
Harun F. Ozbakir;Nolan T Anderson;Kang-Ching Fan;A. Mukherjee
中科院分区：
文献类型：
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作者：
Harun F. Ozbakir;Nolan T Anderson;Kang-Ching Fan;A. Mukherjee
荧光成像是在细胞和分子水平上研究生物功能的基石技术。这项技术的核心是大量收集基于绿色荧光蛋白(GFP)和相关类似物的遗传记者。二十多年的蛋白质工程为GFP谱系提供了令人难以置信的各种荧光蛋白质，使科学家有巨大的自由选择适合各种细胞和环境背景的记者。然而，GFP和衍生品记者有特定的限制，阻碍了它们在分子成像中的不受限制的使用。这些挑战激发了新的报告蛋白和成像机制的发展。在这里，我们回顾了这些发展如何扩展报告基因技术的前沿，使无氧环境和完好动物体内的细胞功能的非破坏性研究成为可能--这两个重要的生物学背景与基于GFP的记者的使用根本不相容。
Fluorescence imaging represents a cornerstone technology for studying biological function at the cellular and molecular levels. The technology's centerpiece is a prolific collection of genetic reporters based on the green fluorescent protein (GFP) and related analogs. More than two decades of protein engineering have endowed the GFP repertoire with an incredible assortment of fluorescent proteins, allowing scientists immense latitude in choosing reporters tailored to various cellular and environmental contexts. Nevertheless, GFP and derivative reporters have specific limitations that hinder their unrestricted use for molecular imaging. These challenges have inspired the development of new reporter proteins and imaging mechanisms. Here, we review how these developments are expanding the frontiers of reporter gene techniques to enable non-destructive studies of cell function in anaerobic environments and deep inside intact animals - two important biological contexts that are fundamentally incompatible with the use of GFP-based reporters.