Generation of T7 RNA Polymerase Transgenic Mouse Strain

利用已完成和正在进行的基因组计划的全部科学潜力，将需要用于创建功能基因组学研究工具的技术进步。由于三种分子在揭示蛋白质功能及其作为新型人类治疗药物的潜力方面发挥着关键作用，因此，制造单克隆抗体的系统和技术的进步尤其至关重要。在本提案中，我们计划创建一个新的系统来衍生单克隆抗体，该系统既简化了开发时间和成本，又专为高通量抗体生产而设计。具体来说，我们计划创造一种表达细胞质RNA聚合酶的转基因小鼠，并将这种小鼠用于阵列产生的线性抗原基因模板的免疫。该系统将消除繁琐和不确定的外源性抗原蛋白生产的需要，并允许同时开发和筛选多种潜在的治疗或诊断相关抗体。在本提案的第一阶段，我们将创建转基因小鼠，开发以阵列形式生产线性抗原基因的技术，并证明该系统使用这些有限的抗原基因集生产抗体的可行性。在第二阶段，我们将在这些小鼠中使用完整的抗原基因阵列来证明该系统如何同时用于筛选多种抗体的功能特性。建议的商业应用：T7RNAP小鼠将成为基因治疗中检测cdna的重要工具，并为杂交瘤的产生提供脾细胞。从事基因治疗研究和单克隆抗体生产的科学家将需要转基因小鼠。这只老鼠可能会被授权给涉及基因治疗的制药公司，以产生特许权使用费收入。我们认为这种鼠标会有很大的商业价值。

Utilizing the full scientific potential of the completed and on-going genome projects will require advances in technologies used to create research tools for functional genomics. Advances in the systems and techniques to create monoclonal antibodies in particular will be crucial importance, due to the critical role thee molecules play in unraveling protein function and their potential as novel human therapeutics. In this proposal, we plan to create a new system to derived monoclonal antibodies, which is both streamlined in development time and costs and is designed for high-throughput antibody production. Specifically, we plan to create a transgenic mouse, which expresses a cytoplasmic RNA polymerase, and use this mouse in immunizations with array-produced linear antigen gene templates. This system will eliminate the need for tedious and uncertain exogenous antigen protein production and allow simultaneous development and screening of multiple potential therapeutically or diagnostically relevant antibodies. In Phase One of this proposal, we shall create the transgenic mouse, develop the technology to produce, in array format, the linear antigen genes, and demonstrate the feasibility of this system to produce antibodies with a limited well- characterized set of these antigen genes. In Phase II, we shall use full arrays of antigen genes in these mice to demonstrate how this system can be used to screen for functional properties of multiple antibodies simultaneously. PROPOSED COMMERCIAL APPLICATIONS: The T7RNAP mouse will be a great tool for testing cDNAs for gene therapy and source of splenocytes for hybridoma production. The transgenic mice would be needed by scientists involved in gene therapy research and monoclonal antibody production. This mouse may be licensed to pharmaceutical companies involve din gene therapy to generate revenues from royalties. We thing this mouse would have great commercial value.