Creating Multi-Gene Reporter Mice Via "Recombineering"

通过“重组工程”创建多基因报告小鼠

• 批准号: 7145213
• 负责人: Alexander C Lichtler
• 金额: $ 14.8万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-17 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7145213
• 关键词: DNA; bone; genes; genetically modified animals; genetics; laboratory mouse; model; motivation; osteoblasts; osteocytes; reporter genes

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.

描述（由申请人提供）：本提案的具体目的是生成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泛的研究应用前景。这将有助于基础科学家了解骨谱系中的细胞是如何生长和分化的。它也可以被私营企业用来帮助鉴定新的化合物，这可能导致治疗骨异常，如骨质疏松症。连锁多基因转基因小鼠的产生是一种新方法，可能影响转基因小鼠的未来设计。我们提出利用细菌人工染色体（BACs）和细菌重组技术组装一个多基因报告基因的DNA结构。将这种转基因小鼠模型与现有小鼠模型区别开来的一个显著特征是，转基因DNA结构将包含多个物理连接的基因报告元件。在同一只小鼠中存在多个报告基因有几个优点，包括提高数据采集率和从同一只小鼠中识别和分离不同细胞群的能力。由于报告基因的连锁，该小鼠模型可以很容易地与其他小鼠系杂交，而无需分离单个报告基因。作为概念的证明，我们选择了三个基因来驱动三种光谱变异荧光报告基因的表达，以鉴定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，我们相信它将有助于改善公共卫生。这将有助于基础科学家了解骨细胞的形成、功能和相互作用。私营企业也可以利用它来帮助鉴定可能导致治疗骨质疏松症等骨骼疾病的新药。