UNS:Plasmonic Nanoantenna-Based Multiplexing microRNA Assay at Zeptomolar Concentrations

UNS：Zeptomolar 浓度下基于等离子纳米天线的多重 microRNA 测定

• 批准号: 1604617
• 负责人: Rajesh Sardar
• 金额: $ 32.91万
• 依托单位: Indiana University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1604617&HistoricalAwards=false
• 关键词: UNS; Plasmonic; Nanoantenna; Based; Multiplexing

PI: Sardar, Rajesh Proposal Number: 1604617MicroRNAs are small nucleic acid strands that are becoming important in understanding fundamental biological pathways and disease diagnostics. The proposed work will attempt measuring them using a technique that is expected to yield very high sensitivity in biological samples.MicroRNAs (miRs) are single-stranded RNAs, capable of regulating genomic processes. Their measurement is useful in disease diagnosis, including cancer. Thus circulating miRs have the potential to serve as diagnostic markers. The current state-of-the-art miR assay is by qRT-PCR, requiring reverse transcription, labeling and amplification. Preliminary studies suggest that hybridizing miR to single-strand DNA (ssDNA) that is attached to gold nanoprisms (GNPs) may induce a novel transduction mechanism via delocalization of conduction electrons. This electronic effect can be investigated utilizing localized surface plasmon resonance (LSPR) properties of GNPs bound to solid substrates. It is found that GNPs undergo a large LSPR wavelength shift upon ssDNA/miR hybridization, and is thought to be due to electronic delocalization, and is an unexplored methodology for analyzing miRs in biological fluids. Further, there is a potential to develop this new methodology into a label-free, low cost multiplexing array. This proposal seeks to validate the electronic delocalization hypothesis by investigating spacing/electronic conjugation between ssDNA and the GNP surface, a single mismatch at different positions in ssDNA/miR helix, chemical functionality attached to ssDNA, and number of nucleotides in miR strands, for structural characterization of the LSPR-based sensors. The proposed research will result in LSPR-based miR sensors capable of assaying at ultralow concentrations in biofluids (Objective 1) via UV-visible spectrometry by monitoring changes in the LSPR peak. To achieve multiplexing capability, portability and long-term stability, GNPs will be immobilized at the bottom of plastic multiwall plates. In this way, each well can be treated as an independent miR sensor (Objective 2). It is expected that assay of 50 distinct miRs excluding positive and negative controls directly from biofluids using a single 96-well plate, and will be characterized in absorption mode by a standard plate reader. Such a design is expected to impact both biomedical research and medical diagnostics. Research activities will involve high school, undergraduate, and graduate students through mentored projects, and the research data will be integrated into existing courses. Nanoscience will be more broadly promoted to middle and high schools through training school teachers in nanostructure synthesis and women in science symposium

PI：Sardar，Rajesh提案编号：1604617 MicroRNAs是一种小的核酸链，在理解基本的生物途径和疾病诊断方面变得越来越重要。这项拟议的工作将尝试使用一种有望在生物样本中产生非常高灵敏度的技术来测量它们。MicroRNAs(MiRs)是单链RNA，能够调节基因组过程。它们的测量在疾病诊断中很有用，包括癌症。因此，循环中的MIR有可能作为诊断标记物。目前最先进的miR检测方法是qRT-PCR，需要逆转录、标记和扩增。初步研究表明，将miR与连接在金纳米颗粒(GNPs)上的单链DNA(SsDNA)杂交，可能通过传导电子的离域作用诱导一种新的转导机制。这种电子效应可以利用结合在固体衬底上的GNPs的局域表面等离子体共振(LSPR)特性来研究。研究发现，GNPs在单链DNA/miR杂交过程中发生了较大的LSPR波长漂移，被认为是由于电子离域引起的，是一种尚未探索的分析生物液中miR的方法。此外，有可能将这种新方法发展成无标签、低成本的多路复用阵列。这一建议试图通过研究单链DNA与GNP表面之间的间距/电子共轭、单链DNA/miR螺旋中不同位置的单个错配、结合在单链DNA上的化学功能以及微链中的核苷酸数量来验证电子离域假说，以用于基于LSPR的传感器的结构表征。这项拟议的研究将导致基于LSPR的miR传感器能够通过监测LSPR峰的变化，在生物体液中以超低浓度进行分析(目标1)。为了实现多路复用能力、便携性和长期稳定性，GNPs将被固定在塑料多壁板的底部。这样，每口井都可以被视为一个独立的MIR传感器(目标2)。预计使用一块96孔板直接从生物体液中检测50种不同的最低抑菌浓度，不包括阳性和阴性对照，并将由标准平板读取器在吸收模式下进行表征。这样的设计预计将对生物医学研究和医疗诊断产生影响。研究活动将通过指导项目涉及高中、本科生和研究生，研究数据将整合到现有课程中。将通过培训学校教师纳米结构合成和妇女科学研讨会，将纳米科学更广泛地推广到初中和高中