Genetically encodable fluorescent biosensors for tracking signaling dynamics in living cells.
Genetically encodable fluorescent biosensors for tracking signaling dynamics in living cells.
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DOI：
10.1021/cr100002u
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发表时间：
2011-05-11
期刊：
影响因子：
62.1
通讯作者：
Zhang, Jin
中科院分区：
文献类型：
--
作者：
Newman, Robert H.;Fosbrink, Matthew D.;Zhang, Jin
荧光一直被认为是探测生物结构和功能的有力工具。大约65年前，Coons和他的同事首次引入荧光抗体技术，作为检测小鼠组织中肺炎球菌抗原的一种手段。[1]在随后的几年里，人们使用免疫荧光和相关技术（如荧光原位杂交）在数百种细胞环境中进行了研究。这些研究为细胞生物学家提供了一个独特的视角来了解细胞和组织的分子结构。然而，由于这种方法通常需要在检测之前进行固定，因此它们仅提供在分子水平上控制细胞生理学的高度动态细胞过程的快照。为了给细胞生物学研究增加一个关键的时间成分，研究人员开发了一套不同的遗传编码生物传感器，旨在以高的空间和时间分辨率探测活细胞中的动态细胞事件。这些传感器通常涉及将荧光标签掺入蛋白质或选定的蛋白质结构域，使研究人员能够在天然细胞环境中真实的时间内跟踪细胞内信号网络的各种组分。在这篇综述中，我们将首先探讨几个基因靶向荧光标记的分子基础，适合活细胞成像。然后，我们将把注意力转向荧光生物传感器的开发和使用，以研究活细胞中的动态信号传导过程。
Fluorescence has long been recognized as a powerful tool for probing biological structure and function. Nearly sixty-five years ago, Coons and colleagues introduced the first fluorescent-antibody techniques as a means of detecting pneumococcal antigens in mouse tissue. 1 In the years that followed, studies have been conducted in hundreds of cellular contexts using immunofluorescence and related techniques such as fluorescence in situ hybridization. 2 Together, these studies have provided cell biologists with a unique glimpse into the molecular organization of cells and tissues. However, because such approaches typically require fixation prior to detection, they provide only a snapshot of the highly dynamic cellular processes that govern cell physiology at the molecular level. To add a crucial temporal component to the study of cell biology, researchers have developed a diverse set of genetically encodable biosensors designed to probe dynamic cellular events in living cells with high spatial and temporal resolution. These sensors, which generally involve the incorporation of a fluorescent tag into a protein or a selected protein domain, have enabled researchers to track various components of intracellular signaling networks in real time within the native cellular environment. In this review, we will first explore the molecular basis of several genetically targetable fluorescent tags amenable to live cell imaging. We will then turn our attention to the development and use of fluorescent biosensors for studying dynamic signaling processes in living cells.
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