A Nanomembrane-Based Nucleic Acid Sensing Platform

1065652常智能优点：便携式和多目标DNA/RNA诊断需要一个快速、现场可用、操作简单、可再生和经济的生物传感器系统。提出的新平台是基于集成了几个新发现的纳米孔膜中的电动现象，有望将传统的无标记电化学和电容/电导生物传感器扩展到所需的稳健性、灵敏度(Pm检测极限约为100万个分子)、选择性(对于kb长的DNA/RNA，在~30个碱基的对接序列上只有一个错配)和检测速度(5分钟)。这些新的检测特征是非线性/选择性离子电导、动态控制的离子耗竭/选择性、浓差极化、分子介电、片上pH控制、电流体动力学、核酸杂交表面电荷反转的结果，所有这些新的纳米孔膜的电动和静电现象都是最近才被理解或发现的，具有最新的纳米制造和成像能力。这项拟议的工作将通过科学地仔细研究详细的非平衡电动现象和利用最新的纳米/微制造技术，将这些新的物理现象优化并集成到适合现场应用的手持DNA/RNA设备的自动膜传感平台中。广泛的影响：拟议的工作将为博士生和博士后提供异常丰富的教育经验。它涉及新的电动和膜物理/化学的基础科学研究、最新的微/纳米制造技术、传染病健康科学、分子遗传学和基因组学、生态学和微型仪器设计，以开发能够对生物研究以及更具商业意义的生物技术工业部门产生重大影响的原型。PI在将团队成员安排到主要研究型大学的终身教职方面有着显著的记录(过去5年中有11个)，其中包括3名NSF职业奖获得者、3名女性和1名非裔美国人。学生和当地高中生/教师也有接受教育的机会，这是通过国际学生联合会实验室的一个积极的暑期推广计划实现的。几乎所有来自PI实验室的本科生研究人员都会继续攻读顶级博士课程。科技影响：一个可行的便携式(手持)和免标签的dna/rna检测平台将推动生物传感器行业的重大技术进步，因为它将从根本上改变医疗、环境、农业和生物防御应用中的病原体检测方法，因为它将从根本上改变医学、环境、农业和生物防御应用中的病原体检测方法。由于几个突出的技术挑战-分析时间慢(比RNA降解时间长)，对样品碎片和化学成分的敏感性，以及昂贵/笨重的检测仪器，健壮和便携的RNA检测技术尚未出现。这项拟议的项目通过互补的基础和制造努力，研究了几种有望缓解这些障碍的新的纳米孔膜现象，并将它们集成到一个多路芯片平台上，可能会导致一种新的分子传感产品。因此，该项目可以对纳米科学和纳米生物技术产生影响。

1065652ChangIntellectual Merit: Portable and multi-target DNA/RNA diagnostics requires a rapid, field-usable, simple to operate, regenerable and economical biosensor system. The proposed new platform is based upon the integration of several newly discovered electrokinetic phenomena in nanoporous membranes that promise to extend traditional label-free electrochemical and capacitance/conductance biosensors to the requisite robustness, sensitivity (pM detection limit of about one million molecules), selectivity (SNP discrimination with only one mismatch at a ~30-base docking sequence for a kb-long DNA/RNA) and assay speed (5 minutes) for on-field nucleic acid detection. These new detection features are results of nonlinear/selective ionic conductance, dynamically controlled ion depletion/selectivity, concentration polarization, molecular dielectrophoresis, on-chip pH control, electrohydrodynamics, surface charge inversion by nucleic acid hybridization all new electrokinetic and electr static phenomena of nanoporous membranes that are only recently understood or discovered with the latest nanofabrication and imaging capabilities. The proposed work will optimize and integrate these new physical phenomena into an automated membrane sensing platform for hand-held DNA/RNA devices suitable for field applications by scientifically scrutinizing the detailed non-equilibrium electrokinetic phenomena and by exploiting the latest nano/microfabrication technologies.Broader Impacts: The proposed work will provide PhD and post-doc students with an unusually rich educational experience. It involves fundamental scientific studies of new electrokinetic and membrane physics/chemistry, the latest micro/nano-fabrication technologies, contagious disease health science,molecular genetics and genomics, ecology and miniature instrumentation design to develop prototypes that can have a significant impact on biological research and, more commercially, the biotechnology industry sector. The PI has significant track records of placing group members in tenure-track faculty positions (11 in the last 5 years) at major research universities, including 3 NSF Career Awardees, 3 women and 1 African American. Educational opportunities also extend to undergraduates and local high school students/teachers through an active summer outreach program in the PI's laboratory. Almost all of the undergraduate researchers from the PI's lab go on to top PhD programs. Significant opportunities exist for international collaborations due to the PI's strong ties to institutions in Taiwan, Korea, Europe, and China through his capacity as the founding and chief editor of Biomicrofluidics, an American Institute of Physics journal with high impact factor.Scientific and Technological Impact: A viable portable (handheld) and label-free DNA/RNA detection platform for viral assays, bacteria detection etc. will spur a major technological advance in the biosensor industry, as it will fundamentally transform pathogen detection methodology for medical, environmental, agricultural and biodefense applications by eliminating the time-consuming PCR or reverse transcription PCR step and the costly/bulky/personnel-intensive optical detectors of fluorescent sensing platforms. Robust and portable RNA detection technologies have yet to appear because of several outstanding technological challenges---slow assay time (longer than RNA degradation time), sensitivity to sample debris and chemical content, and expensive/bulky detection instrumentation. The proposed project investigates, with complementary fundamental and fabrication efforts, several new nanoporous membrane phenomena that promise to alleviate these obstacles and integrates them onto a multiplex chip platform that may lead to a new molecular sensing product. The project can hence impact both nano science and nano biotechnology.