I-Corps: Gel-Tethered Molecular Beacons

I-Corps：凝胶束缚分子信标

• 批准号: 1262903
• 负责人: Matthew Libera
• 金额: $ 5万
• 依托单位: Stevens Institute of Technology
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-10-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1262903&HistoricalAwards=false
• 关键词: Corps; Gel; Tethered; Molecular; Beacons

Molecular Beacons are self-reporting DNA hairpin probes that have been used in a variety of real-time detection applications. When they hybridize to an oligonucleotide target they fluoresce, thus removing the additional requirement of fluorescently labeling the target itself. Consequently, molecular beacons have found substantial scientific success over the past two decades. Molecular beacons have rarely been used in microarray-based applications. When tethered to solid substrates a large background emerges due to beacon-substrate interactions. This large background is absent when beacons are used un-tethered in an aqueous solution. Researchers have discovered that the signal-to-background ratio of molecular beacons can remain relatively high if they are tethered to a solid substrate by means of a hydrogel. This high performance has been attributed to the fact that the crosslink density of surface-patterned microgels gradually approaches infinity at the microgel-water interface, and the beacons conjugated to the microgel surface find themselves in as water-like an environment as possible. This project is designed to demonstrate the effective use of surface-tethered molecular beacons in clinically relevant DNA assays. Since their invention in the mid-1990's, molecular beacons and beacon-based assay platforms have served multiple scientific and commercial roles. However, one large applications area where beacons have rarely been used is in microarray technology where the oligonucleotide probes must be tethered to a solid surface. Site-specific surface tethering is important, because the nature of a specific probe and its target can be identified by position rather than by fluorophore color. Thousands of discrete surface positions can be interrogated simultaneously whereas the practical number of different fluorophores is less than ten. Applications involving the rapid identification of bacterial strains responsible for a broad range of different infections, the ability to simultaneously probe for tens or hundreds of genes can have substantial impact in health-care settings has the potential to be enhanced through the results of this project.

分子信标是自我报告的DNA发夹探针，已用于各种实时检测应用。当它们与寡核苷酸目标杂交时，它们发出荧光，从而消除了对目标本身进行荧光标记的额外要求。因此，分子信标在过去二十年中取得了重大的科学成功。分子信标很少用于基于微阵列的应用。当连接到固体衬底时，由于信标-衬底相互作用，会出现一个大背景。当信标在水溶液中使用时，不存在这种大背景。研究人员发现，如果通过水凝胶将分子信标固定在固体基质上，它们的信本比可以保持相对较高的水平。这种高性能归因于这样一个事实，即表面图案微凝胶的交联密度在微凝胶-水界面处逐渐接近无穷大，并且与微凝胶表面共轭的信标发现自己处于尽可能像水的环境中。该项目旨在展示在临床相关的DNA分析中有效使用表面栓系分子信标。自20世纪90年代中期发明以来，分子信标和基于信标的分析平台已经发挥了多种科学和商业作用。然而，信标很少使用的一个大的应用领域是微阵列技术，其中寡核苷酸探针必须拴在固体表面。位点特异性表面系固很重要，因为可以通过位置而不是荧光团颜色来识别特定探针及其目标的性质。数千个离散的表面位置可以同时被询问，而不同荧光团的实际数量少于10个。涉及快速识别导致各种不同感染的细菌菌株的应用，同时探测数十或数百个基因的能力可以在保健环境中产生重大影响，有可能通过这个项目的结果得到加强。