Tools for live cell imaging of biomolecular interactions

生物分子相互作用的活细胞成像工具

• 批准号: 298496-2010
• 负责人: Pezacki, JohnPaul
• 金额: $ 3.64万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=571503
• 关键词: Tools; live; cell; imaging; biomolecular

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. This research program will offer training opportunities for graduate students in chemistry to work at the interface between chemistry and biology, recieve multidisciplinary training that is so desperately needed for the competitiveness of Canadian biotech sector, emerging areas of academic and governmental research. Trainees from my lab have acquired positions in major pharmaceutical companies such as Roche and Amgen and PhD students have gone on to post-doctoral positions with scholarship support to prestigious institututions such as UBC and Stanford.

灵敏的分子成像技术代表了诊断学的未来。需要进行基础研究，以开发有助于可视化疾病状态的分子探针，结合纳米级分辨率成像技术，可以在细胞阶段检测疾病的分子决定因素，通常是在给定组织中疾病的表型明显之前。在这项发现补助金中，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，用于可视化致病性RNA，并可视化细胞膜上的宿主-病毒相互作用。这两种技术将用于可视化细胞和组织中的丙型肝炎病毒（HCV）RNA，以及导致HCV细胞进入的纳米级相互作用的图像。虽然这些工具和方法可以更广泛地应用，但我们选择在HCV上展示它们，因为它是一种慢性感染超过25万加拿大人的病毒，而且很难检测到。将合成新型和高灵敏度的探针，用于感染细胞中丙型肝炎病毒RNA的直接成像。 将探讨一些探针设计。一旦构建探针，将进行HCV感染细胞的体内成像。我们还建议对不同的探针结构进行基础研究，以研究使用我们实验室开发的纳米级成像技术在细胞进入过程中涉及的分子过程。 该研究计划将为化学研究生提供培训机会，在化学和生物学之间的接口工作，接受加拿大生物技术部门竞争力迫切需要的多学科培训，学术和政府研究的新兴领域。我实验室的实习生已经在罗氏和安进等大型制药公司获得了职位，博士生已经在卑诗大学和斯坦福大学等著名机构获得奖学金支持的博士后职位。