I-Corps: Gel-Tethered Molecular Beacons

分子发夹是自报告DNA发夹探针，已用于各种实时检测应用。当它们与寡核苷酸靶杂交时，它们发出荧光，从而消除了荧光标记靶本身的额外要求。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。当拴系到固体基板时，由于信标-基板相互作用而出现大的背景。当信标在水溶液中不受约束地使用时，不存在这种大的背景。研究人员已经发现，如果分子信标通过水凝胶拴在固体基质上，它们的信号与背景比可以保持相对较高。 这种高性能归因于表面图案化的微凝胶的交联密度在微凝胶-水界面处逐渐接近无穷大，并且缀合至微凝胶表面的信标发现它们自己处于尽可能类似水的环境中。该项目旨在证明表面拴系分子信标在临床相关DNA检测中的有效使用。自20世纪90年代中期发明以来，分子信标和基于信标的测定平台已经发挥了多种科学和商业作用。然而，很少使用信标的一个大的应用领域是在微阵列技术中，其中寡核苷酸探针必须拴系到固体表面。位点特异性表面束缚是重要的，因为特异性探针及其靶标的性质可以通过位置而不是通过荧光团颜色来识别。可以同时询问数千个离散的表面位置，而不同荧光团的实际数量小于10个。涉及快速鉴定导致广泛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能对医疗保健环境产生重大影响，这些应用有可能通过该项目的结果得到加强。

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.