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

点击翻译按钮获取中文摘要

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.