NEW METHOD FOR DIRECT AND QUANTITATIVE RNA DETECTION

直接定量 RNA 检测的新方法

• 批准号: 2644167
• 负责人: MARY A BROW
• 金额: $ 10万
• 依托单位: THIRD WAVE TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2644167
• 关键词: RNA; biotechnology; gene expression; measurement; method development; nuclease; oligonucleotides; protein engineering

DESCRIPTION (Adapted from investigator's abstract): The goal of this Phase I proposal is the development of a simple, cost-effective, and high-throughput method for direct and quantitative RNA detection. In molecular medicine, such a method would greatly facilitate the analysis of RNA viruses and the measurement of specific gene expression, currently pressing problems in the field. Despite this need, few techniques have emerged that are truly direct and quantitative over a broad range of concentration differences, while being readily adaptable to any genetic system. This proposal describes the development of a novel enzymatic assay for direct and quantitative RNA analysis in a microtiter plate-based format. This assay relies on structure-specific cleavage of oligonucleotides hybridized to the target RNA and can be used to detect any sequence. A combination of protein engineering and solid-phase capture approaches will be used to attain a sensitivity of detection of subattomole quantities of RNA without intervening conversion to DNA. The expected outcome of Phase I is an assay for quantitative monitoring of gene expression in 1 mg or less of tissue that will be adaptable to any RNA sequence. This system will be adaptable to a variety of low-cost readout platforms, including chemiluminescent, fluorescent and colorimetric detection. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE

描述(改编自调查人员摘要)：本报告的目标 第一阶段的建议是开发一种简单、经济、 高通量直接定量检测RNA的方法。在……里面 分子医学，这样的方法将极大地方便分析 RNA病毒和特定基因表达的测量， 目前这一领域的紧迫问题。尽管有这种需求，但很少有人 已经出现了真正直接和定量的技术 广泛的浓度差异，同时易于适应 任何遗传系统。本提案描述了一种 一种新的直接定量分析细菌RNA的酶促方法 基于微滴定板的格式。这种分析依赖于结构特异性 与靶RNA杂交的寡核苷酸的切割可以 用于检测任何序列。蛋白质工程和蛋白质工程的结合 将使用固相捕获方法来获得以下灵敏度 无干预检测核糖核酸亚摩尔量 转化为DNA。第一阶段的预期结果是一种检测 定量监测1毫克或更少组织中的基因表达 这将适用于任何RNA序列。这个系统将是 可适应各种低成本读出平台，包括 化学发光、荧光和比色检测。 建议的商业应用：不可用