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

PI：Sardar，Rajesh 提案编号：1604617微小RNA是小的核酸链，在理解基本生物学途径和疾病诊断中变得越来越重要。 这项工作将尝试使用一种有望在生物样品中产生非常高灵敏度的技术来测量它们。microRNAs（miRs）是能够调节基因组过程的单链RNA。它们的测量可用于疾病诊断，包括癌症。因此，循环miR具有用作诊断标志物的潜力。目前最先进的miR测定是通过qRT-PCR，需要逆转录、标记和扩增。初步研究表明，将miR与连接到金纳米棱镜（GNP）的单链DNA（ssDNA）杂交可能会通过传导电子的离域诱导一种新的转导机制。这种电子效应可以利用GNP与固体基质结合的局部表面等离子体共振（LSPR）性质来研究。发现GNP在ssDNA/miR杂交后经历大的LSPR波长偏移，并且被认为是由于电子离域，并且是用于分析生物流体中的miR的未探索的方法。此外，有可能将这种新方法发展成无标记、低成本的多路复用阵列。该提案旨在通过研究ssDNA和GNP表面之间的间隔/电子缀合、ssDNA/miR螺旋中不同位置处的单个错配、附着于ssDNA的化学功能性和miR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。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，通过紫外可见光谱法在生物流体中的超低浓度下进行测定（目标1）。为了实现多路复用能力、便携性和长期稳定性，GNP将被固定在塑料多壁板的底部。以这种方式，每个孔可以被视为独立的miR传感器（目的2）。预期使用单个96孔板直接从生物流体中测定50种不同的miR（不包括阳性和阴性对照），并将通过标准酶标仪以吸收模式进行表征。这种设计预计将影响生物医学研究和医学诊断。研究活动将涉及高中，本科和研究生通过指导项目，研究数据将被整合到现有的课程。纳米科学将通过培训学校教师纳米结构合成和妇女参与科学研讨会更广泛地推广到初中和高中

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