Plasmon Coupling Microscopy for the in vivo Detection and Tracking of Cytoplasmic

描述(申请人提供)：本方案描述了一种新的成像技术，该技术可以实时检测和跟踪活细胞中的单个RNA分子，而不受观察时间的限制。这项新技术是基于单个贵金属纳米颗粒之间的距离依赖等离子激元耦合，这些纳米颗粒被RNA探针链拴住。互补RNA靶链的杂交会引起颗粒间距离的变化，从而导致等离子体共振波长的移动。这种光谱漂移作为指示靶向RNA结合的信任点。等离子体激元耦合方法的特殊新奇之处在于所提出的纳米粒子传感器的主动性质。由于它们固有的相互耦合能力，所提出的传感器能够连续监测RNA系绳的端到端距离。所提出的传感器不仅可以检测和跟踪单个RNA分子，还可以实时提供有关标记的RNA系链的结构变化的信息。该提案的具体目标是：目标1：开发RNA偶联的贵金属纳米二聚体，用于体内应用。目的2：建立基于等离子激元耦合的单个RNA分子检测技术。目的3：实时检测和跟踪活细胞中的单个胞质RNA分子。这项拟议的研究对健康的好处包括，有可能开发出有效的方法来沉默导致癌症等严重疾病的基因。生物医学界的其他好处包括新技术，它允许检测活细胞中的单个RNA分子，从而允许在单细胞水平上进行具有未知灵敏度的全新诊断。

DESCRIPTION (provided by applicant): This proposal describes a new imaging technology that can detect and track single RNA molecules in living cells in real time without limitation in observation time. The new technology is based on the distance dependent plasmon coupling between individual noble metal nanoparticles that are tethered by a RNA probe strand. Hybridization of a complementary RNA target strand induces an interparticle distance change that leads to a shift in the plasmon resonance wavelength. This spectral shift serves as fiduciary point indicating target RNA binding. The special novelty of the plasmon coupling approach is the active nature of the proposed nanoparticle sensors. Due to their inherent ability to couple with each other, the proposed sensors enable a continuous monitoring of the end-to-end distance of the RNA tether. The proposed sensors cannot only detect and track individual RNA molecules, they will also provide information about structural changes of the labeled RNA tether in real-time. The specific aims of the proposal are: Aim 1: Develop RNA coupled noble metal nanoparticle dimers for in vivo applications. Aim 2: Develop a plasmon coupling based detection technology for individual RNA molecules. Aim 3: Detect and track individual cytoplasmic RNA molecules in living cells in real time. The health benefits of the proposed research include the potential to develop effective ways to silence genes that cause serious diseases such as cancer. Other benefits to the biomedical community include new technologies that allow the detection of single RNA molecules in living cells thereby allowing an entirely new diagnostics with unknown sensitivity on the single cell level.