Department of Bioengineering, Center for Biophysics and Computational Biology, Institute of Genomic Biology, The Beckman Institute for Advance Science and Technology, University of Illinois at Urbana-Champaign, IL 61801, USA.
Integr Biol (Camb). 2009 Oct;1(10):565-73. doi: 10.1039/b913093b.
Since the development of green fluorescent protein (GFP) and other fluorescent proteins (FPs) with distinct colors, genetically-encoded probes and biosensors have been widely applied to visualize the molecular localization and activities in live cells. In particular, biosensors based on fluorescence resonance energy transfer (FRET) have significantly advanced our understanding of the dynamic molecular hierarchy at subcellular levels. These biosensors have also been extensively applied in recent years to study how cells perceive the mechanical environment and transmit it into intracellular molecular signals (i.e. mechanotransduction). In this review, we will first provide a brief introduction of the recent development of FPs. Different FRET biosensors based on FPs will then be described. The last part of the review will be dedicated to the introduction of examples applying FRET biosensors to visualize mechanotransduction in live cells. In summary, the integration of FRET technology and the different cutting-edge mechanical stimulation systems can provide powerful tools to allow the elucidation of the mechanisms regulating mechanobiology at cellular and molecular levels in normal and pathophysiological conditions.
自从绿色荧光蛋白 (GFP) 和其他具有独特颜色的荧光蛋白 (FPs) 的发展以来,遗传编码探针和生物传感器已广泛应用于可视化活细胞中的分子定位和活性。特别是基于荧光共振能量转移 (FRET) 的生物传感器极大地促进了我们对亚细胞水平动态分子层次结构的理解。近年来,这些生物传感器也被广泛应用于研究细胞如何感知机械环境并将其转化为细胞内分子信号(即机械转导)。在这篇综述中,我们将首先简要介绍 FPs 的最新发展。然后将描述基于 FPs 的不同 FRET 生物传感器。综述的最后一部分将专门介绍应用 FRET 生物传感器可视化活细胞中机械转导的实例。总之,将 FRET 技术与不同的尖端机械刺激系统相结合,可以为阐明正常和病理生理条件下细胞和分子水平上调节机械生物学的机制提供强大的工具。