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.

1149015-当前分析挑战的复杂性增加需要新颖，有效的分离和纯化技术。例如，分析各种生物分子需要强大的分离和纯化技术，这些通常是生化研究中的限制成分，例如生物标志物发现，临床诊断或单细胞分析。新型大分子结构（例如人造DNA纳米组装）的分离和纯化对于成功的纳米技术应用，例如对于DNA计算，在光子设备中或目标诊断中，这进一步至关重要。该提案由化学，生物工程，环境和运输系统划分的化学和生物学分离计划支持，化学分工的化学测量和成像计划支持DNA介电嗜介摄取（DEP）及其在浓度，分数和分离DNA的应用方面的研究。现象介电性电泳是指在不均匀电场中可极化分子的迁移。不同DNA物种极化的变化将导致迁移的差异，这可以用于分析目的。然而，迄今为止，DNA的极化机制仍然几乎无法理解，这阻碍了介电的使用用于分析应用。因此，该提案的目的是针对各种DNA的极化性的定量研究，不仅允许将介电水素应用于分离和浓缩，而且还揭示了DNA极化性的起源。使用基于绝缘体的DEP的量身定制的小型平台将用于建立必要的电场条件以使DEP发生。采用的微流体平台进一步提供了DNA的微型和快速分析，并将允许分析适用于小细胞组合甚至单个细胞中DNA的pico-至纳米含量范围的微小样品。将研究DNA的传输机制，以揭示其与DNA DEP的相互作用与优化DNA分析的最终目标。基于这些知识，提出了从DNA纳米技术对DNA疫苗的质量控制到基于DNA的诊断的分析应用。该项目还提出了针对ASU的化学和生物化学系的女性本科和研究生化学学生的指导计划。拟议的活动旨在鼓励和促进妇女，这些活动在更高职业阶段的化学方面仍然不足。该计划包括个人指导活动以及针对女性本科生的一般活动，包括与本提案的智力优点有关的研究机会。在研究生层面，该计划专门增强了这些指导活动，以减轻沟通，科学表现和谈判技巧以及网络机会，博士学位进步和职业发展。