Li Isaac T, Pham Elizabeth, Truong Kevin
Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ont., Canada, M5S 3G9.
Biotechnol Lett. 2006 Dec;28(24):1971-82. doi: 10.1007/s10529-006-9193-5. Epub 2006 Oct 5.
Genetically-coded, fluorescence resonance energy transfer (FRET) biosensors are widely used to study molecular events from single cells to whole organisms. They are unique among biosensors because of their spontaneous fluorescence and targeting specificity to both organelles and tissues. In this review, we discuss the theoretical basis of FRET with a focus on key parameters responsible for designing FRET biosensors that have the highest sensitivity. Next, we discuss recent applications that are grouped into four common biosensor design patterns--intermolecular FRET, intramolecular FRET, FRET from substrate cleavage and FRET using multiple colour fluorescent proteins. Lastly, we discuss recent progress in creating fluorescent proteins suitable for FRET purposes. Together these advances in the development of FRET biosensors are beginning to unravel the interconnected and intricate signalling processes as they are occurring in living cells and organisms.
基因编码的荧光共振能量转移（FRET）生物传感器被广泛用于研究从单细胞到整个生物体的分子事件。由于其自发荧光以及对细胞器和组织的靶向特异性，它们在生物传感器中独具特色。在本综述中，我们讨论FRET的理论基础，重点关注负责设计具有最高灵敏度的FRET生物传感器的关键参数。接下来，我们讨论近期的应用，这些应用被分为四种常见的生物传感器设计模式——分子间FRET、分子内FRET、底物切割产生的FRET以及使用多种颜色荧光蛋白的FRET。最后，我们讨论在创建适用于FRET目的的荧光蛋白方面的最新进展。FRET生物传感器开发中的这些进展共同开始揭示活细胞和生物体中正在发生的相互关联且错综复杂的信号传导过程。
