NIRT: DNA Sequencing and Translocation Studies using Electrically-Addressable Nanopore Arrays

本提案是对NSF 03-043 NIRT类纳米科学与工程倡议的响应。该研究计划将利用来自不同学科的PI和Co-PI的免费专业知识来开发新一代固态纳米孔设备，该设备将能够以前所未有的精度进行测量，并探索新的DNA测序机制。NIRT团队将：(1)开发一种电可寻址纳米孔阵列(EANA)设备及其相关的微流体系统。(2)开发利用EANA和生物纳米孔的混合纳米孔-纳米孔系统，并开展首次多路复用溶血素纳米孔实验；(3)探索利用孔区多肽核酸(PNA)分子功能化的SiO_2纳米孔进行序列检测的可行性；(4)对共同PI提出的使用单链结合蛋白驱动DNA易位的理论模型进行实验测试，并考察其对测序的影响；(5)开展利用EANA技术在单分子水平实时监测DNA降解过程的可行性研究，及其在DNA测序方面的潜力。这些NIRT活动将在生物和生物医学研究中产生广泛的影响，为研究生物分子提供新的技术和更有效的分析工具。这些项目为处于纳米科学、单分子生物物理和理论物理前沿的博士后和研究生提供了极好的培训机会。该项目还为布朗大学和哈佛大学的本科生提供了新的令人兴奋的纳米生物科学跨学科领域的研究经验。这些学生将接受硅纳米制造、纳米和微流体、光学、分子生物物理和多体统计物理方面的高级培训。这些新技能将为学生未来在工业、学术界和公共服务领域的职业生涯做好准备。该奖项的资金由数学和物理科学局的物理和材料科学司、工程局的工程教育和中心司以及生物科学局的生物基础设施司提供。

This proposal was received in response to Nanoscale Science and Engineering initiative, NSF 03-043, category NIRT. The research program will utilize the complimentary expertise of the PI and co-PIs from different disciplines to develop a new generation of solid-state nanopore device that will allow measurements with unprecedented accuracies, and exploration of new DNA sequencing mechanisms. The NIRT team will: (1) Develop an electrically-addressable nanopore array (EANA) device and its related microfluid systems. (2) Develop a hybrid nanopore-in-nanopore system using EANA and biological nanopores, and carry out the first multiplexing Hemolysin nanopore experiments; (3) Explore the feasibility of sequence detection using SiO2 nanopores functionalized with polypeptide nucleic acid (PNA) molecules in the pore region; (4) Carry out an experimental test of a theoretical model proposed by a co-PI using single strand binding proteins to drive DNA translocation, and to examine its implications for sequencing; (5) Carry out a feasibility study of using the EANA technology for real-time monitoring of DNA degradation processes at single molecule level, and its potential for DNA sequencing. These NIRT activities will have broad impact in biological and biomedical research, in providing new techniques and more efficient analytical tools for studying biomolecules. The projects provide excellent training opportunities for postdoctoral and graduate students at the forefront of nanoscience and single-molecule biophysics and theoretical physics. The program also offers undergraduate students at Brown University and Harvard University new exciting research experience in the interdisciplinary field of nanobioscience. The students will receive advanced training in silicon nanofabrication, nano- and microfluidics, optics, molecular biophysics, and many-body statistical physics. These new skills will prepare the students for future careers in industry, academia, and public service. Funds for this award are provided by the Divisions of Physics and Materials Science in the Mathematical and Physical Sciences Directorate, the Division of Engineering Education and Centers in the Engineering Directorate, and the Division of Biological Infrastructure in the Directorate for Biological Sciences.