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.

该奖项支持预期能够检测出低丰度（Zeptomoles或更少）生物分子靶标的超敏感光谱传感系统的开发，而无需扩增。 该系统将能够在单个平台上对DNA，RNA和蛋白质进行多重分析。 目前，使用PCR进行目标扩增是检测低丰度核酸的最有效方法。但是，PCR倾向于诱导背景放大，这通常会破坏其准确性。 由于无法获得蛋白质的扩增技术，因此很难检测到低含量。综合基因组和蛋白质组学分析的越来越重要，例如mRNA和蛋白质表达的相关测量值需要在单个平台上进行基因组和蛋白质组学分析的通用生物分析工具。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整合。 该系统将由能够检测核酸和蛋白质靶标的通用纳米传感器以及能够多路复用的单分子荧光检测的4色微流体SMD光谱仪组成。 纳米传感器将由功能化的量子点（QD）组成，该点将产生结合诱导的荧光共振能传递（FRET）信号。 当与4色微流体SMD光谱仪结合使用时，如果不放大，则可以对低通量靶标进行高通量检测。 该设备的能力将通过单个细胞中模型基因表达系统的mRNA和蛋白表达的同时测量，以及通过未膨胀基因组DNA中的单核苷酸多态性（SNP）的基因分型来证明。 这项工作的跨学科性质将为研究生和本科阶段的跨学科培训提供绝佳的机会。 成功开发了微流体设备后，学生将参与一系列独立研究项目，该项目在生物元，微/纳米流体和纳米生物传感器系统中。 预计高中生还将参与该项目的某些方面。