IDBR: Development of a Universal Spectroscopic Nanosensing System for Multiplexed Genomic and Proteomic Analysis

IDBR：开发用于多重基因组和蛋白质组分析的通用光谱纳米传感系统

• 批准号: 0552063
• 负责人: Jeff Wang
• 金额: $ 30万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-15 至 2008-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0552063&HistoricalAwards=false
• 关键词: IDBR; Development; Universal; Spectroscopic; Nanosensing

This award supports development of an ultrasensitive spectroscopic sensing system expected to be able to detect low-abundance (zeptomoles or less) biomolecular targets without the need for amplification. This system will be capable of multiplexed analysis of DNA, RNA, and protein on a single platform. Target amplification using PCR is currently the most effective approach for detection of low-abundance nucleic acids; however, PCR tends to induce background amplification which often undermines its accuracy. Because an amplification technique for proteins is not available, detection of low-abundance is difficult. The increasing importance of integrated genomic and proteomic analysis, e.g. correlated measurements of mRNA and protein expression, calls for a universal bioanalytical tool capable of both genomic and proteomic analysis on a single platform. The sensing system will be based on innovative integration of single-molecule detection (SMD), micro/nanofluidics, and molecular biology. The system will be comprised of a generic nanosensor capable of detecting both nucleic acids and protein targets and a 4-color microfluidic SMD spectroscope capable of multiplexed fluorescence detection of single molecules. The nanosensor will consist of functionalized quantum dots (QDs) that will give a binding-induced fluorescence resonance energy transfer (FRET) signal. When combined with the 4-color microfluidic SMD spectroscope, high-throughput detection of low-abundance targets will be possible without amplification. The capability of the device will be demonstrated through simultaneous measurements of mRNA and protein expression in a model gene expression system in single cells, and through genotyping single-nucleotide polymorphisms (SNPs) in unamplified genomic DNA. The interdisciplinary nature of the effort will provide excellent opportunities for cross-disciplinary training at the graduate and undergraduate level. After successful development of the microfluidic devices, students will be involved in a series of independent research projects in the areas of BioMEMS, micro/nanofluidics, and nanobiosensor systems. High school students are also expected to participate in some aspects of the project.

该奖项支持开发一种超灵敏的光谱传感系统，该系统有望在不需要放大的情况下检测低丰度(Zeptomoles或更少)生物分子目标。该系统将能够在单一平台上对DNA、RNA和蛋白质进行多路分析。靶向扩增技术是目前检测低丰度核酸最有效的方法，然而，扩增往往会导致背景扩增，这往往会影响其准确性。因为没有蛋白质的放大技术，所以很难检测到低丰度。基因组和蛋白质组综合分析的重要性日益增加，例如对信使核糖核酸和蛋白质表达的相关测量，这就需要一种能够在单一平台上同时进行基因组和蛋白质组分析的通用生物分析工具。该传感系统将基于单分子检测(SMD)、微/纳米流体和分子生物学的创新集成。该系统将由一个能够同时检测核酸和蛋白质靶标的通用纳米传感器和一个能够对单分子进行多路荧光检测的四色微流控SMD光谱仪组成。该纳米传感器将由功能化量子点(QD)组成，这些量子点将发出结合诱导的荧光共振能量转移(FRET)信号。当与四色微流控SMD光谱仪结合使用时，无需放大即可实现低丰度目标的高通量检测。该设备的能力将通过同时测量单个细胞中模型基因表达系统中的mRNA和蛋白质表达，以及通过对未扩增基因组DNA中的单核苷酸多态(SNPs)进行基因分型来证明。这项工作的跨学科性质将为研究生和本科生一级的跨学科培训提供极好的机会。在成功开发出微流控设备后，学生将参与生物微机械、微/纳米流体学和纳米生物传感器系统等领域的一系列独立研究项目。预计高中生也将参与该项目的某些方面。