Creating Multi-Gene Reporter Mice Via "Recombineering"

描述(申请人提供)：本方案的具体目标是建立一种与骨生物学相关的物理连接的多基因报告小鼠模型。该动物模型在骨生物学领域具有广阔的研究应用前景。它将帮助基础科学家了解骨系中的细胞如何生长和分化。它也可能被私营企业用来帮助鉴定新的化合物，这可能导致治疗骨骼异常，如骨质疏松症。连锁多基因转基因小鼠的产生是新的，可能会影响未来转基因小鼠的设计。我们建议使用细菌人工染色体(BAC)和细菌内重组技术来组装一个多基因报告DNA结构。将这种转基因小鼠模型与目前存在的模型区分开来的一个明显特征是，转基因DNA结构将包含多个物理上相连的基因报告元件。在同一只小鼠中存在多个报告基因有几个优点，包括提高数据获取的速度，以及从同一只小鼠中识别和分离不同的细胞群的能力。由于报告基因的连锁，这种小鼠模型可以很容易地与其他品系的小鼠杂交，而不需要单独分离记者。作为概念的证明，我们选择了三个基因来驱动三个光谱变异荧光报告的表达，以识别对骨生物学重要的三种不同的细胞类型。这种动物模型将在研究中有广泛的应用，我们相信它将为改善公共卫生做出贡献。它将帮助基础科学家了解骨细胞如何形成、功能和相互作用。它还可能被私营企业用来帮助识别新药，这些新药可能会导致治疗骨质疏松等骨骼疾病。

DESCRIPTION (provided by applicant): The specific aim of this proposal is to generate a physically linked multiple-gene-reporter mouse model that is relevant to bone biology. This animal model will have broad research applications in the field of bone biology. It will aid basic scientists in their ability to understand how cells within the bone lineage grow and differentiate. It may also be used by private industry to aid in the identification of novel compounds, which may lead to treatment of bone abnormalities, such as osteoporosis. The generation of linked multi-gene transgenic mice is novel and may influence the future design of transgenic mice. We propose to assemble a multi-gene-reporter DNA construct using Bacteria Artificial Chromosomes (BACs) and recombination technology in bacteria. A distinct feature that separates this transgenic mouse model from currently existing ones, is that the transgenic DNA construct will contain multiple gene reporter elements that are physically linked. The presence of multiple reporter genes in the same mouse has several advantages, including increasing the rate of data acquisition and the ability to identify and isolate distinct cell populations from the same mouse. As a result of linkage of the reporter genes, this mouse model can be easily crossed with other lines of mice without separation of individual reporters. As proof of concept, we have chosen three genes to drive the expression of three spectral variant fluorescent reporters to identify three distinct cell types important to bone biology. This animal model will have broad applications in research and we believe it will contribute to improvements in public health. It will aid basic scientists in their ability to understand how bone cells form, function, and interact with each other. It may also be used by private industry to aid in the identification of new drugs, which may lead to treament of bone diseases, such as osteoporosis.