Next Generation Plasmon Coupling Nanosensors

下一代等离子耦合纳米传感器

• 批准号: 2344525
• 负责人: Bjoern Reinhard
• 金额: $ 44.77万
• 依托单位: Trustees of Boston University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2344525&HistoricalAwards=false
• 关键词: Next; Generation; Plasmon; Coupling; Nanosensors

Ribonucleic acid (RNA) molecules have diverse functions in controlling gene expression and controlling cellular behavior. RNA is also used by some viruses to store genetic information. These diverse and important functions motivate the development of sensors that allow for a sensitive detection and identification of RNA molecules. This project will develop RNA sensors for two specific classes of RNA molecules: i.) microRNA (miRNA), a regulatory class of RNA that has potential as biomarkers for diseases such as cancer, and ii.) viral RNA to enable a rapid detection of RNA viruses, such as the corona virus (Covid-19). The proposed sensor platform contains a DNA strand connected at its ends to two metal nanoparticles whose optical signal depend on their separation. The DNA sensor strand is designed to change its structure when it binds to a specific RNA target. This structural modification changes the separation between the nanoparticle probes and alters their signal, which is sensitively detected even for small nanoparticles via interference of the scattered signal with a reference light beam. The sensor will be designed to detect miRNA directly, but for viral RNA the coupled nanoparticle sensor will be combined with an enzymatic signal amplification step to obtain a sensitive and rapidly reconfigurable detection platform. The research will be integrated into education and outreach activities that include development of course work to support the research, organization of a summer workshop on the fundamentals of plasmonics and nanoscience for inner city high school students in the Boston area and their teachers that is designed to excite them about a career in science and technology, and development of an Instagram channel that will provide an excellent platform for educating a lay audience about plasmonic nanosensors.The project will combine molecular beacon like DNA structures whose ends are functionalized with noble metal nanoparticle (NP) probes with two-color interferometric scattering microscopy (iSCAT) detection to generate a plasmon coupling based sensor for two important classes of RNA: i.) regulatory miRNA and ii.) severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. Target RNA binding to the sensor induces a conformational change from a “closed” to an “open” conformation and alters the plasmon coupling between the NP labels bound to the DNA strand. Two-color iSCAT microscopy will be applied to enhance the sensitivity for detecting changes in the interferometric scattering signal associated with the opening of the sensor upon binding to its target. The DNA recognition element of the sensor will be designed to bind to miRNA directly, while for the viral RNA the plasmonic transducer will be combined with CRISPR-CAS13 detection and subsequent sample amplification to obtain a sensitive and reconfigurable viral RNA detection platform. Instead of directly binding to viral RNA, the molecular beacon will recognize RNA fragments generated through CAS13 trans-cleavage of a substrate.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

核糖核酸 (RNA) 分子在控制基因表达和控制细胞行为方面具有多种功能。一些病毒还使用 RNA 来存储遗传信息。这些多样化且重要的功能推动了传感器的开发，以实现对 RNA 分子的灵敏检测和识别。该项目将为两类特定的 RNA 分子开发 RNA 传感器：i.) microRNA (miRNA)，一种调节性 RNA，有潜力作为癌症等疾病的生物标志物；ii.) 病毒 RNA，能够快速检测冠状病毒 (Covid-19) 等 RNA 病毒。所提出的传感器平台包含一条 DNA 链，其末端连接到两个金属纳米颗粒，其光学信号取决于它们的分离。 DNA 传感器链被设计为在与特定 RNA 靶标结合时改变其结构。这种结构修饰改变了纳米颗粒探针之间的间距并改变了它们的信号，即使对于小纳米颗粒，也可以通过散射信号与参考光束的干涉来灵敏地检测到信号。该传感器将被设计用于直接检测 miRNA，但对于病毒 RNA，耦合的纳米颗粒传感器将与酶信号放大步骤相结合，以获得灵敏且快速可重构的检测平台。该研究将被纳入教育和外展活动中，包括开发课程以支持研究、为波士顿地区内城高中生及其老师组织关于等离子体和纳米科学基础知识的夏季研讨会，旨在激发他们对科学技术职业的兴趣，以及开发 Instagram 频道，为非专业观众提供有关等离子体纳米传感器的绝佳平台。该项目将结合分子信标等 DNA 结构的末端用贵金属纳米粒子 (NP) 探针进行功能化，并进行双色干涉散射显微镜 (iSCAT) 检测，以生成基于等离激元耦合的传感器，用于检测两类重要的 RNA：i.) 调节性 miRNA 和 ii.) 严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) RNA。目标 RNA 与传感器的结合诱导构象从“闭合”构象变为“开放”构象，并改变与 DNA 链结合的 NP 标记之间的等离激元耦合。双色 iSCAT 显微镜将用于增强检测与传感器结合目标后打开相关的干涉散射信号变化的灵敏度。传感器的DNA识别元件将被设计为直接与miRNA结合，而对于病毒RNA，等离子体传感器将与CRISPR-CAS13检测和随后的样本扩增相结合，以获得灵敏且可重构的病毒RNA检测平台。该分子信标不会直接与病毒 RNA 结合，而是识别通过 CAS13 底物反式切割生成的 RNA 片段。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。