CAREER: DNA Analysis Based on Dielectrophoresis

职业：基于介电泳的 DNA 分析

• 批准号: 1149015
• 负责人: Alexandra Ros
• 金额: $ 49.4万
• 依托单位: Arizona State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-15 至 2018-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1149015&HistoricalAwards=false
• 关键词: CAREER; DNA; Analysis; Based; Dielectrophoresis

1149015 - RosThe increasing complexity of current analytical challenges demands novel and efficient separation and purification techniques. For example, analyzing a large variety of biomolecules requires powerful separation and purification techniques, and these are often the limiting component in biochemical research such as in biomarker discovery, clinical diagnosis or single cell analysis. The separation and purification of novel macromolecular structures such as artificial DNA nano-assemblies is further essential for their successful nano-technological applications, such as for DNA computing, in photonic devices or in targeted diagnostics. This proposal supported by the Chemical and Biological Separations Program of the Chemical, Bioengineering, Environmental and Transport Systems Division and the Chemical Measurement and Imaging Program of the Chemistry Division supports aims in the study of dielectrophoresis (DEP) of DNA and its application for concentrating, fractionating and separating DNA. The phenomenon dielectrophoresis refers to the migration of polarizable molecules in inhomogeneous electric fields. Variations in the polarization of different DNA species will lead to differences in migration, which can be exploited for analytical purposes. To date however, the polarization mechanism of DNA remains only little understood, which hampers the use of dielectrophoresis for analytical applications. This proposal thus aims at a quantitative study of the polarizability for a wide variety of DNA not only allowing the application of dielectrophoresis for separation and pre-concentration but also to reveal the origin of DNA polarizability. A tailored miniaturized platform employing insulator-based DEP will be used to establish the necessary electric field conditions for DEP to occur. The employed microfluidic platforms further provide miniaturized and fast analysis of DNA by DEP, and will allow the analysis of minute samples in the range of pico- to nano-liters suitable for the analysis of DNA in small cell ensembles or even single cells. Concomitantly occurring transport mechanisms for DNA will be investigated to reveal their interplay with DNA DEP with the ultimate goal to optimize DNA analysis. Based on this knowledge, analytical applications ranging from DNA nanotechnology over quality control of DNA vaccines to DNA-based diagnostics are proposed.This project also proposes a mentoring plan for female undergraduate and graduate chemistry students in the Chemistry and Biochemistry Department at ASU. The proposed activities aim in encouragement and promotion of women, which are still underrepresented in chemistry at higher career stages. The plan includes individual mentoring activities as well as general activities for female undergraduate students including research opportunities related to the intellectual merit of this proposal. At the graduate level, the plan specifically intensifies these mentoring activities to ameliorate communication, scientific presentation and negotiation skills as well as networking opportunities, PhD progress and career development.

当前日益复杂的分析挑战需要新颖高效的分离和纯化技术。例如，分析各种各样的生物分子需要强大的分离和纯化技术，而这些通常是生物化学研究（如生物标志物发现、临床诊断或单细胞分析）中的限制因素。新型大分子结构的分离和纯化，如人工DNA纳米组件，对于其成功的纳米技术应用，如DNA计算、光子设备或靶向诊断，是必不可少的。该提案由化学、生物工程、环境和运输系统部门的化学和生物分离计划以及化学部门的化学测量和成像计划支持，旨在研究DNA的介电电泳（DEP）及其在DNA浓缩、分离和分离中的应用。介质电泳现象是指极化分子在不均匀电场中的迁移。不同DNA物种的极化变化将导致迁移的差异，这可以用于分析目的。然而，到目前为止，DNA的极化机制仍然知之甚少，这阻碍了使用介电电泳的分析应用。因此，本提案旨在定量研究多种DNA的极化性，不仅允许应用介电分离和预浓缩，而且揭示DNA极化性的起源。采用基于绝缘子的DEP的定制小型化平台将用于建立发生DEP所需的电场条件。所采用的微流控平台进一步通过DEP提供小型化和快速的DNA分析，并将允许分析微米至纳米升范围内的微小样品，适用于分析小细胞群甚至单细胞中的DNA。同时发生的DNA转运机制将被研究，以揭示它们与DNA DEP的相互作用，最终目标是优化DNA分析。基于这一知识，提出了从DNA纳米技术到DNA疫苗质量控制到基于DNA的诊断的分析应用。本项目还为亚利桑那州立大学化学与生物化学系的女本科生和研究生提出了一个指导计划。拟议的活动旨在鼓励和促进妇女，因为在较高职业阶段从事化学工作的妇女人数仍然不足。该计划包括个别指导活动以及为女本科生提供的一般活动，包括与该提案的智力价值相关的研究机会。在研究生阶段，该计划特别加强了这些指导活动，以改善沟通，科学演讲和谈判技能以及网络机会，博士学位的进步和职业发展。