Generation of T7 RNA Polymerase Transgenic Mouse Strain

利用已完成和正在进行的基因组项目的全部科学潜力，将需要在用于创建功能基因组学研究工具的技术方面取得进展。由于这些分子在解开蛋白质功能和它们作为新的人类疗法的潜力中所起的关键作用，创造单抗的系统和技术的进步将特别重要。在这项提案中，我们计划创建一种新的获得单抗的系统，该系统在开发时间和成本方面都得到了简化，并设计用于高通量抗体生产。具体地说，我们计划创造一种表达细胞质RNA聚合酶的转基因小鼠，并将其用于阵列生产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。在这项计划的第一阶段，我们将创造转基因小鼠，开发以阵列形式生产线性抗原基因的技术，并证明该系统生产具有有限的、特征良好的这些抗原基因的抗体的可行性。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演示如何使用该系统同时筛选多种抗体的功能特性。拟议的商业应用：T7RNAP小鼠将是测试用于基因治疗的cDNA的伟大工具，以及用于杂交瘤生产的脾细胞的来源。参与基因治疗研究和克隆抗体生产的科学家将需要转基因小鼠。这种小鼠可能会被授权给涉及DIN基因治疗的制药公司，以从版税中获得收入。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.