Unified Reporter Gene for Multi-Modality Imaging

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。它通常涉及用报告基因测定真实的实时监测活细胞和组织中的基因转录和调控。一个可以在活细胞和动物中工作的报告基因将是非常有价值的，但目前还没有。本研究的总体目标是开发一种整合的报告基因系统，该系统适用于单个活细胞和包括人类在内的整个活动物中的基因表达成像。 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。为了将这种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基板成像转基因表达与正电子发射断层扫描（PET）在整个生活的小鼠。这些新的探针将在具有肿瘤异种移植物的活小鼠中进行验证。将评估其灵敏度和特异性，并与现有的PET成像报告基因测定进行比较。 这项工作将提供一个统一的报告基因系统，可以监测基因表达从单一的活细胞进行高通量选择整个活的动物与PET。这种多功能的系统在生物学中具有广泛的应用，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。

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.