OPTICALLY ENHANCED OLIGONUCLEOTIDES TO SCREEN FOR METABOLIC DISEASES

用于筛选代谢疾病的光学增强寡核苷酸

• 批准号: 8360021
• 负责人: ANDREA E HOLMES
• 金额: $ 4.23万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360021
• 关键词: Address; Binding; Biological Assay; Color; Complex; Development; Devices; Disease Marker; Dyes; Fluorescence; Fluorescence Spectroscopy; Fluorescent Dyes; Funding; Gel; Grant; Maps; Mass Screening; Measures; Medical; Metabolic Diseases; Methods; Molecular; Monitor; National Center for Research Resources; Nebraska; Newborn Infant; Oligonucleotides; Polymers; Principal Investigator; Property; Research; Research Infrastructure; Resources; Screening procedure; Silicates; Solid; Source; Steroids; Testing; Time; United States National Institutes of Health; Urine; aptamer; base; combinatorial; cost; cost effective; functional genomics; interest; molecular recognition; sensor; steroid metabolism

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Current screening methods for metabolic diseases are too costly, expensive, and time-consuming for broadly testing all newborns. Thus, many metabolic diseases go undetected and untreated for many years and screening is implemented only for the one most common metabolic disease. Using the proposed sol-gel supported aptamer-dye mix-and-measure assays in urine allows for cost-effective mass screening. The roposed research is dedicated to the development of new, highly selective, and sensitive molecular sensors that change color or fluorescence properties in the presence of steroid metabolites. Several steroids are disease markers for inborn metabolic diseases. We are interested in mapping three-way junctions of oligonucleotide-based binders (aptamers) to determine general principles that can be used in understanding molecular recognition of fluorescent dyes, visible dyes, steroids, and their metabolites by aptamers. The screening of aptamer-dye complexes will address an important medical need by generating cost-effective, rapid, mix-and-measure methods suitable for mass screening and monitoring of steroids or metabolites caused by faulty steroid metabolism. Organic dyes that form strong complexes with binding aptamers and metabolites will be found using a combinatorial 96-well plate approach. Reliable aptamer binding assays by UV-Vis and fluorescence spectroscopy have been established by our group. Furthermore, aptamers will be incorporated into colloidal silicate polymers for the development of handheld solid sensor devices to be used in medicinal applications. "Sol-gels" are based on solid colloidal silicates that will form a porous hard gel and will incorporate the aptamer-dye complexes within the gel, allowing for attaching the new molecular sensors into a solid support.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 目前的代谢性疾病筛查方法过于昂贵、昂贵和耗时，无法对所有新生儿进行广泛的测试。因此，许多代谢性疾病多年来没有被发现和治疗，只对一种最常见的代谢性疾病进行筛查。使用建议的溶胶-凝胶支持的适体染料混合和测量尿液检测方法，可以进行经济高效的大规模筛查。这项研究致力于开发新的、高选择性和灵敏的分子传感器，在类固醇代谢物存在的情况下改变颜色或荧光性质。几种类固醇是先天代谢性疾病的疾病标志。我们感兴趣的是绘制基于寡核苷酸的结合体(适配子)的三向连接，以确定可用于理解适配子对荧光染料、可见染料、类固醇及其代谢物的分子识别的一般原理。适体-染料复合体的筛选将通过产生经济高效、快速、混合和测量的方法来满足重要的医疗需求，适用于大规模筛选和监测由类固醇代谢故障引起的类固醇或代谢物。有机染料与结合适体和代谢物形成强大的络合物将被发现使用组合96孔板方法。本课题组建立了可靠的紫外-可见光谱和荧光光谱适体结合分析方法。此外，适体将被加入到胶体硅酸盐聚合物中，以开发用于医疗应用的手持式固体传感器设备。“溶胶-凝胶”是以固体胶体硅酸盐为基础的，这种固体胶体硅酸盐将形成一种多孔的硬凝胶，并将适体-染料复合体结合到凝胶中，从而将新的分子传感器连接到固体载体中。