Biosensing in electrochemically controlled nano-junctions

电化学控制纳米结中的生物传感

• 批准号: 8257908
• 负责人: Krzysztof Slowinski
• 金额: $ 10.73万
• 依托单位: CALIFORNIA STATE UNIVERSITY LONG BEACH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8257908
• 关键词: Binding; Biosensing Techniques; Buffers; Complex; DNA; DNA-Binding Proteins; Data; Detection; Development; Diagnosis; Electric Conductivity; Electrochemistry; Electrodes; Electron Transport; Fluorescence; Funding; Future; Goals; Gold; Grant; Image; Individual; Intercalating Agents; Laboratories; Length; Light; Link; Literature; Measures; Medicine; Methods; Microscopy; Miniaturization; Molecular; Monitor; Optics; Point Mutation; Point-of-Care Systems; Positioning Attribute; Process; Productivity; Protein Binding; Protocols documentation; Reading; Research; Research Infrastructure; Research Project Grants; Resolution; Scanning; Scanning Tunneling Microscopy; Scheme; Science; Scientist; Sequence-Specific DNA Binding Protein; Single Nucleotide Polymorphism; Solutions; Spottings; Students; TATA-Box Binding Protein; Technology; Training; United States National Institutes of Health; aqueous; base; biochip; cost effective; electrical property; graduate student; light intensity; nano; nanodevice; nanometer; nanosensors; programs; public health relevance; single molecule; undergraduate student

DESCRIPTION (provided by applicant): The progress in medicine is inherently linked to the development of robust technologies for the detection of biomolecules. The biochip technology provides accurate, practical, and cost-effective systems for point of care diagnosis via quantification of the molecular complexes using colorimetric, fluorescence, or electrochemical methods. The future development of the biochip methods depends on further miniaturization. It is clear, furthermore, that the processing of readout from biochips based on light intensity is fundamentally limited by the relationship between the size of the individual spot and the wave-length of light. This places a limitation on the accuracy of optical readout from the nanochips and thus provides an argument for further development of electrical detection schemes. Such electrical detection should enable the biochip technology to break through into the nanometer region since the resolution of the readings would be determined, in principle, by the size of a single DNA molecule. The goal of this proposal is to develop experimental strategies for the electrical detection of single-nucleotide polymorphisms (SNPs) and sequence- specific DNA-binding proteins (e.g. TATA binding protein) at the single molecule level via monitoring of the electrical properties of DNA molecules modified with covalently attached Nile Blue (NB) in electrochemically controlled nanoscopic tunnel junctions. The key objective of the proposed studies is to determine whether disruption of the DNA -stack via a single-point mutation or a protein binding results in an analytically effectual decrease in electrical conductivity of the helix. Specifically, we plan to develop protocols for the imaging of NB modified DNA arrays with single-molecule resolution via electrochemically controlled scanning tunneling microscopy, EC-STM. We will also use EC-STM approach to detect single nucleotide polymorphisms and to detect TATA binding protein (TBP) at the single molecule level. Finally, we will use the electrical conductivity of the NB modified DNA helix interposed between two closely spaced gold electrodes as an analytical nano-device for the detection of TBP based on sequence-specific binding to DNA. The PI's developmental objective is to continue engagement in bio-medically related research projects. It is expect that the requested SC-3 support would increase PI's productivity and would allow PI to continue research program with graduate and undergraduate students at CSULB. The completion of a four year funding requested in this application should allow PI to become more competitive for major NIH/NSF grant support. This funding will ultimately support the research infrastructure sustaining PI's efforts to entice students with the excitement of science and to train them to become our future scientists and professionals. PUBLIC HEALTH RELEVANCE: The biochip technology provides accurate, practical, and cost-effective systems for point of care diagnosis. The future development of the biochip methods depends on further miniaturization. The goal of this proposal is to develop experimental strategies for the electrical detection of single-nucleotide polymorphisms (SNPs) and sequence-specific DNA-binding proteins (e.g. TATA binding protein) at the single molecule level. Demonstrating the feasibility of such detection is critically important in the development of DNA based nano-sensors.

描述（由申请人提供）：医学的进度与生物分子检测的强大技术的开发固有地联系在一起。生物芯片技术通过使用比色法，荧光或电化学方法定量分子复合物，提供了准确，实用和成本效益的系统，用于诊断。生物芯片方法的未来发展取决于进一步的微型化。此外，很明显，基于光强度的生物芯片的读数从根本上受到了从单个点的大小与光长度之间的关系的限制。这限制了纳米芯片光学读数的准确性，因此为进一步开发电检测方案提供了一个论点。这种电气检测应使生物芯片技术能够通过单个DNA分子的大小确定读数的分辨率，从而使生物芯片技术突破到纳米区域。 该提案的目的是开发实验策略，以通过在单分子水平上监测DNA分子的DNA分子的电型nile蓝色（NBLECH）nile蓝色（NB）中的电型蓝色（NB）中的电气级别的电位，以在单分子水平上监测单分子水平的单分子水平上的单分子级别电位（例如TATA结合蛋白）的单个核苷酸多态性（SNP）和序列 - 特异性DNA结合蛋白（例如TATA结合蛋白）。连接。拟议研究的主要目的是确定通过单点突变或蛋白质结合对DNA堆的破坏是否会导致螺旋电导率的分析有效降低。具体而言，我们计划通过电化学控制的扫描隧道显微镜EC-STM来开发具有单分子分辨率的NB修饰DNA阵列成像的协议。我们还将使用EC-STM方法检测单个核苷酸多态性并在单分子水平上检测TATA结合蛋白（TBP）。最后，我们将使用插入两个紧密间隔的金电极之间的NB修饰的DNA螺旋的电导率作为分析纳米装置，以基于序列特异性结合与DNA的结合来检测TBP。 PI的发展目标是继续参与生物医学相关的研究项目。预计要求的SC-3支持将提高PI的生产率，并允许PI继续与CSULB的研究生和本科生一起研究计划。在本申请中要求的四年资金的完成应使PI在NIH/NSF主要赠款支持方面变得更有竞争力。这笔资金最终将支持维持PI努力的研究基础设施，以吸引学生兴奋，并训练他们成为我们未来的科学家和专业人士。 公共卫生相关性：生物芯片技术为护理点诊断提供了准确，实用且具有成本效益的系统。生物芯片方法的未来发展取决于进一步的微型化。该提案的目的是开发实验策略，以在单分子水平上进行单核苷酸多态性（SNP）和序列特异性DNA结合蛋白（例如TATA结合蛋白）的电气检测。证明这种检测的可行性对于基于DNA的纳米传感器的发展至关重要。