Artificial Nanotube Gels for Mutation Detection

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 为特定疾病提供无创、低成本、快速筛查的技术 癌症的分子特征。高度敏感和具体，DOVAM-S (几乎所有突变的检测-SSCP)能够筛选出几个 人类基因突变的准确率几乎为100%。实现的关键技术 展示了一种用于检测的纳米颗粒、电泳式微器件 几乎所有类型的突变和多态。有了这种技术， 具有成本效益的癌症易感性筛查可以在 相对近期而言。基于碳纳米管的独特性质，我们 设想一种试剂显著减少的纳米DOVAM-S一体化体系 体积、缩短的放大和分离时间以及自动读出 以及对结果的解释。在这份提案中，我们提出了这一愿景，并 进行概念验证演示的必要步骤：首先证明 碳纳米管阵列的SSCP是可能的，随后是协议 临床突变检测标准DOVAM-S的研制与比较 筛选实验。开发芯片上DNA分析的其他努力 技术缺乏区别于DOVAM-S的本质优势 相互竞争的突变筛查方法。这几乎是100% 检测率和多路采样的能力。为了有效地发挥作用， 用DOVAM-S进行突变筛查需要对单个突变进行电泳法分离 带有和不带有突变的搁浅DNA。我们的方法是使用碳 纳米管作为一种固态纳米级电泳筛的研究 单链DNA的分离，取代了过去使用的交联聚合物 常规凝胶电泳法。相互竞争的努力提供芯片上的DNA 分析保留了交联型聚合物的使用，许多分析是基于 杂交反应，不能100%检测突变 这一点在这里倡导的DOVAM-S方法中得到了证明。纳米DOVAM- S将使用人类因子和IX基因以及自动取款机基因进行验证， 在某些癌症中发生突变。

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.