Carbon Nanotubes: Artifical Gels for Mutations Detection

碳纳米管：用于突变检测的人造凝胶

• 批准号: 6440122
• 负责人: STEVE Seev SOMMER
• 金额: $ 16.28万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-17 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6440122
• 关键词: DNA; atomic force microscopy; biomedical automation; buffers; cancer risk; carbon; charge coupled device camera; chemical synthesis; computer data analysis; conformation; diagnosis design /evaluation; diagnostic tests; electrochemistry; electrophoresis; gene mutation; genetic disorder diagnosis; genetic polymorphism; genetic screening; intermolecular interaction; nanotechnology; nucleic acid purification; nucleic acid sequence; scanning electron microscopy; serine proteinases; technology /technique development; water

DESCRIPTION (provided by applicant): Our vision is to combine nanotechnology with proven genetic screening technology to provide non-invasive, low cost, rapid screening for specific molecular signatures of cancer. Highly sensitive and specific, DOVAM-S (Detection of Virtually All Mutations-SSCP) is capable of screening several human genes for mutations with virtually 100% accuracy. The key technology to be demonstrated is a nanofabricated, electrophoretic microdevice for detecting mutations and polymorphisms of virtually all types. With this technology, cost-effective screening for cancer predisposition could be implemented in the relatively near term. Based on the unique properties of carbon nanotubes, we envision an integrated nano-DOVAM-S system with significantly reduced reagent volumes, shortened amplification and separation times, and automated readout and interpretation of results. In this proposal, we present this vision, and the necessary steps toward a proof-of-concept demonstration: first proving that SSCP is possible with carbon nanotube arrays, followed by protocol development and comparison with standard DOVAM-S in clinical mutation screening experiments. Other efforts to develop on-chip DNA analysis technology lack the essential advantages that distinguish DOVAM-S from competing approaches to mutation screening. These are a virtually 100% detection rate and the ability to multiplex samples. In order to be effective, mutation screening by DOVAM-S requires electrophoretic separation of single- stranded DNA with and without mutations. Our approach is to use carbon nariotubes as a solid-state nanometer-scale sieve for electrophoretic separation of single-strand DNA, replacing the cross-linked polymers used in conventional gel electrophoresis. Competing efforts to provide on-chip DNA analysis retain the use of cross-linked polymers, and many are based on hybridization reactions, which cannot attain the 100% detection of mutations that has been demonstrated in the DOVAM-S approach advocated here. Nano-DOVAM- S will be validated using the human factor VIII and IX genes and the ATM gene, mutated in certain cancers.

描述（由申请人提供）： 我们的愿景是将纳米技术与经过验证的基因筛查相结合 技术为特定人群提供非侵入性、低成本、快速筛查 癌症的分子特征。高度灵敏和特异，DOVAM-S （检测几乎所有突变-SSCP）能够筛选几种 人类基因突变的准确度几乎为 100%。关键技术 被证明是一种纳米制造的电泳微型器件，用于检测 几乎所有类型的突变和多态性。有了这项技术， 可以在以下地区实施具有成本效益的癌症易感性筛查： 相对近期。基于碳纳米管的独特性能，我们 设想一种显着减少试剂的集成纳米 DOVAM-S 系统 体积、缩短的扩增和分离时间以及自动读数 和结果的解释。在本提案中，我们提出了这一愿景，并且 概念验证演示的必要步骤：首先证明 SSCP 可以通过碳纳米管阵列实现，然后是协议 临床突变中标准 DOVAM-S 的开发和比较 筛选实验。开发芯片 DNA 分析的其他努力 技术缺乏区分 DOVAM-S 的本质优势 突变筛选的竞争方法。这些几乎是 100% 检测率和多重样本的能力。为了有效， DOVAM-S 突变筛选需要电泳分离单突变 有或没有突变的链DNA。我们的方法是利用碳 nariotubes 作为用于电泳的固态纳米级筛子 单链DNA的分离，取代了用于的交联聚合物 常规凝胶电泳。提供芯片 DNA 的竞争努力 分析保留了交联聚合物的使用，并且许多基于 杂交反应，无法达到100%突变检测 这里提倡的 DOVAM-S 方法已经证明了这一点。纳米DOVAM- S 将使用人类因子 VIII 和 IX 基因以及 ATM 基因进行验证， 在某些癌症中发生突变。