Tools for live cell imaging of biomolecular interactions

灵敏的分子成像技术代表着诊断学的未来。需要基础研究来开发有助于可视化疾病状态的分子探针，结合纳米分辨率成像技术可以在细胞阶段检测疾病的分子决定因素，通常在给定组织中疾病的表型明显之前。在这项发现拨款中，我们建议对不同探针设计的化学和生物学进行基础研究，以开发和研究新的分子成像工具，用于可视化致病RNA和可视化细胞膜上的宿主与病毒的相互作用。这两项技术将被应用于可视化细胞和组织中的丙型肝炎病毒(HCV)RNA，以及导致丙型肝炎病毒进入细胞的纳米级相互作用的图像。尽管这些工具和方法可能会得到更广泛的应用，但我们选择在丙型肝炎病毒上进行演示，因为它是一种慢性感染超过25万加拿大人的病毒，很难检测到。将合成新的、高灵敏度的探针，用于直接成像感染细胞中的丙型肝炎病毒RNA。将探索一些探头设计。一旦构建了探针，将对感染丙型肝炎病毒的细胞进行体内成像。我们还建议对不同的探针结构进行基础研究，以利用我们实验室正在开发的纳米级成像技术来研究细胞进入过程中涉及的分子过程。

Sensitive molecular imaging techniques represent the future of diagnostics. Fundamental research is needed to develop molecular probes that help to visualize disease states combined with nanoscale resolution imaging techniques allows for the detection of molecular determinants of disease at the cellular stage often before the phenotype of disease is apparent in a given tissue. In this discovery grant we propose to perform fundamental research into the chemistry and biology of different probe designs in order to develop and investigate new molecular imaging tools for the visualization of pathogenic RNA and to visualize host-virus interactions at the cellular membrane. These two techniques will be applied to visualize hepatitis C virus (HCV) RNA in cells and tissues, as well as image at the nanoscale interactions that lead to HCV cell entry. Although the tools and approaches may be applied more broadly we have chosen to demonstrate them on HCV because it is a virus that chronically infects more than 250,000 Canadians and it is difficult to detect. Novel and highly sensitive probes will be synthesized for the direct imaging of hepatitis C virus RNA in infected cells. A number of probe designs will be explored. Once the probes are constructed, in vivo imaging of HCV-infected cells will be conducted. We also propose to perform fundamental studies on different probe architectures for the purposes of investigating molecular processes involved during cell entry processes using nanoscale imaging techniques being developed in our lab.