Programmable Multi-Target Detection Using an Aptamer-Integrated Nanopore

使用适配体集成纳米孔进行可编程多目标检测

基本信息

批准号：
8018612
负责人：
Li-Qun (Andrew) GU
金额：
$ 24.91万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8018612
关键词：
Address Affinity Aptamer Technology Area Binding Biological Process Biomedical Engineering Biomedical Technology Biotechnology Cell physiology Cells Chemical Engineering Complex DNA DNA Fingerprinting Detection Dimensions Disease Dissection Engineering Environmental Engineering technology Frequencies Gene Expression Generations Goals HIV-1 IgE Influenza Ions Laboratories Lead Life Ligands Light Mediating Modeling Molecular Nanotechnology Nucleic Acids Osmosis Property Proteins RNA RNA-Directed DNA Polymerase Reagent Research Science Solutions Surface Technology Testing Viral aptamer base biological systems chemical reaction design detector high throughput screening improved molecular scale nano nanobiotechnology nanofabrication nanomedicine nanopore nanoscale novel therapeutics particle pathogen protein protein interaction receptor single molecule success tool voltage

项目摘要

DESCRIPTION (provided by applicant): This proposal addresses an interdisciplinary strategy that integrates the use of highly sensitive, highly specific aptamers with nanopore-based single molecule technology. The long term goal is to develop aptamer-integrated nanopore probes for biomedical detections. Aptamers are engineered DNA/RNA that can specifically recognize broad species of proteins with high affinities. Upon binding, these powerful molecules can inhibit pathogen protein, catalyze chemical reactions, control gene expression, and regulate cellular functions. Therefore aptamers can potentially be applied as tools for exploring biological systems. Motivated by such small, but sophisticated molecules, we would like to integrate aptamer technology with our nanopore probe to construct a new generation of single molecule detector that would aid greatly in diverse disease-related detections. Nanopore technology can "visually" capture the dynamic binding of a single molecule to a ligand in a nanometer-scaled pore through the discrete changes in conductance upon binding. We propose the combined use of nanopore with laboratory nanofabrication, bio-friendly surface engineering, and molecular engineering to accomplish the following specific aims: 1. Understand and manipulate various properties of nanopores for improvement of single molecule detection; 2. Establish a fundamental model for single molecule detection with aptamer-equipped nanopores. Broad target species with varied aptamer types (DNA/RNA) will be tested, including Immunoglobulin E, HIV-1 reverse transcriptase, and large influenza A viral particle; 3. Establishing an advanced model that employs engineered Transfer Aptamer (t-Aptamer) to mediate detection. The t-Aptamer, which composes an aptamer and a universal carrier, acts as a "translator" to encode target information into the frequency of hybridization between the carrier and the immobilized receptor in the nanopore. Such a t-Aptamer-mediated detection is robust, programmable, and will ultimately lead to a universal nanopore that can discriminate different targets and perform simultaneous multi-target detection. The success of this research will greatly expand the capability of nanopores as the new generation of detection technology for biomedical analysis and high-throughput screening. Fashioning such tools is significant in nanomedicine for the quantitative characterization and precise control of molecular scale components (or nanomachinery) of living cells.

描述（由申请人提供）：该提案提出了一种跨学科策略，将高灵敏度、高特异性适体的使用与基于纳米孔的单分子技术相结合。长期目标是开发用于生物医学检测的适配体集成纳米孔探针。适配体是经过改造的 DNA/RNA，可以特异性识别具有高亲和力的多种蛋白质。结合后，这些强大的分子可以抑制病原体蛋白、催化化学反应、控制基因表达并调节细胞功能。因此，适体可以作为探索生物系统的工具。受到如此小而复杂的分子的启发，我们希望将适配体技术与我们的纳米孔探针相结合，构建新一代单分子检测器，这将极大地帮助各种疾病相关的检测。纳米孔技术可以通过结合时电导的离散变化“直观地”捕获单个分子与纳米级孔中配体的动态结合。我们建议将纳米孔与实验室纳米制造、生物友好的表面工程和分子工程相结合，以实现以下具体目标： 1. 理解和操纵纳米孔的各种特性，以改进单分子检测； 2. 建立适配体纳米孔单分子检测的基本模型。将测试具有不同适配体类型（DNA/RNA）的广泛目标物种，包括免疫球蛋白E、HIV-1逆转录酶和大甲型流感病毒颗粒； 3. 建立利用工程转移适体（t-适体）介导检测的先进模型。 t-Aptamer由适配体和通用载体组成，充当“翻译器”，将目标信息编码为载体与纳米孔中固定化受体之间的杂交频率。这种t适体介导的检测是稳健的、可编程的，并且最终将产生一种通用的纳米孔，可以区分不同的目标并同时执行多目标检测。这项研究的成功将极大扩展纳米孔作为新一代生物医学分析和高通量筛选检测技术的能力。制造此类工具对于纳米医学中活细胞分子级组件（或纳米机械）的定量表征和精确控制具有重要意义。