A Unified Reporter Gene for Multi-Modality Imaging

多模态成像的统一报告基因

• 批准号: 6909071
• 负责人: Jianghong Rao
• 金额: $ 24万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6909071
• 关键词: athymic mouse; autoradiography; beta lactamase; bioimaging /biomedical imaging; biotechnology; carbon; chemical synthesis; fluorine; gene expression; high throughput technology; laboratory mouse; molecular probes; nanotechnology; positron emission tomography; radionuclides; reporter genes; single cell analysis; tissue /cell culture; transfection; xenotransplantation

DESCRIPTION (provided by applicant): Understanding gene function is a fundamental but still challenging task. It often involves monitoring gene transcription and regulation in both live cells and tissues in real time with reporter gene assays. A reporter gene that can work in both live cells and animals would be very valuable, but is currently unavailable. The overall goals of this research are to develop an integrated reporter gene system that is suitable for imaging gene expression in both single living cells and whole living animals including humans. An ampicillin resistance gene (encoding a bacterial enzyme, beta-lactamase) is an outstanding reporter for optical measurement of gene expression in single living mammalian tissue culture cells. To extend this reporter gene to whole living animals, it will require the development of new substrates. We will design and synthesize new substrates for imaging transgene expression with positron emission tomography (PET) in whole living mice. These new probes will be validated in living mice with tumor xenografts. Their sensitivity and specificity will be assessed and compared to the existing PET imaging reporter gene assays. This work will provide a unified reporter gene system that can monitor gene expression from single live cells undergoing high-throughput selection to whole living animals with PET. Such a versatile system should find wide applications in biology from in vivo cell trafficking, tumor metastasis, small animal disease models, transgenic mice, drug screenings in mice, to in vivo gene function and regulation study.

描述（由申请人提供）： 了解基因功能是一项基本但仍然具有挑战性的任务。它通常涉及通过报告基因检测实时监测活细胞和组织中的基因转录和调控。能够在活细胞和动物中发挥作用的报告基因将非常有价值，但目前还无法获得。这项研究的总体目标是开发一个集成的报告基因系统，适用于单个活细胞和包括人类在内的整个活体动物的基因表达成像。 氨苄青霉素抗性基因（编码细菌酶β-内酰胺酶）是用于光学测量单个活哺乳动物组织培养细胞中基因表达的杰出报告基因。为了将该报告基因扩展到整个活体动物，需要开发新的底物。我们将设计和合成新的底物，用于在活体小鼠中使用正电子发射断层扫描（PET）对转基因表达进行成像。这些新探针将在带有肿瘤异种移植物的活体小鼠中进行验证。他们的敏感性和特异性将被评估，并与现有的 PET 成像报告基因检测进行比较。 这项工作将提供一个统一的报告基因系统，可以利用 PET 监测从经过高通量选择的单个活细胞到整个活体动物的基因表达。这种多功能系统应该在生物学中得到广泛的应用，从体内细胞运输、肿瘤转移、小动物疾病模型、转基因小鼠、小鼠药物筛选到体内基因功能和调控研究。