Hochreiter Bernhard, Garcia Alan Pardo, Schmid Johannes A
Institute for Vascular Biology and Thrombosis Research, Medical University Vienna, Schwarzspanierstraße17, Vienna A-1090, Austria.
Sensors (Basel). 2015 Oct 16;15(10):26281-314. doi: 10.3390/s151026281.
Fluorescence- or Förster resonance energy transfer (FRET) is a measurable physical energy transfer phenomenon between appropriate chromophores, when they are in sufficient proximity, usually within 10 nm. This feature has made them incredibly useful tools for many biomedical studies on molecular interactions. Furthermore, this principle is increasingly exploited for the design of biosensors, where two chromophores are linked with a sensory domain controlling their distance and thus the degree of FRET. The versatility of these FRET-biosensors made it possible to assess a vast amount of biological variables in a fast and standardized manner, allowing not only high-throughput studies but also sub-cellular measurements of biological processes. In this review, we aim at giving an overview over the recent advances in genetically encoded, fluorescent-protein based FRET-biosensors, as these represent the largest and most vividly growing group of FRET-based sensors. For easy understanding, we are grouping them into four categories, depending on their molecular mechanism. These are based on: (a) cleavage; (b) conformational-change; (c) mechanical force and (d) changes in the micro-environment. We also address the many issues and considerations that come with the development of FRET-based biosensors, as well as the possibilities that are available to measure them.
荧光或Förster共振能量转移（FRET）是一种可测量的物理能量转移现象，发生在合适的发色团之间，当它们足够接近时，通常在10纳米范围内。这一特性使它们成为许多关于分子相互作用的生物医学研究中极其有用的工具。此外，这一原理越来越多地被用于生物传感器的设计，其中两个发色团与一个控制它们距离从而控制FRET程度的传感域相连。这些FRET生物传感器的多功能性使得以快速和标准化的方式评估大量生物变量成为可能，不仅允许进行高通量研究，还能对生物过程进行亚细胞测量。在本综述中，我们旨在概述基于荧光蛋白的基因编码FRET生物传感器的最新进展，因为这些代表了基于FRET的传感器中最大且增长最活跃的群体。为便于理解，我们根据其分子机制将它们分为四类。这些类别基于：（a）切割；（b）构象变化；（c）机械力；（d）微环境变化。我们还讨论了基于FRET的生物传感器开发过程中出现的许多问题和注意事项，以及测量它们的可用方法。
