Collal Driven Ligand-Regulated Cre Transgenic Mice

Collal 驱动的配体调节 Cre 转基因小鼠

• 批准号: 6442053
• 负责人: Alexander C Lichtler
• 金额: $ 7.21万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-28 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6442053
• 关键词: beta galactosidase; biotechnology; bone metabolism; developmental genetics; drug administration rate /duration; gene expression; genetically modified animals; green fluorescent proteins; laboratory mouse; ligands; osteoblasts; recombinase; reporter genes; tamoxifen; technology /technique development; transfection /expression vector

DESCRIPTION (provided by applicant): The goal of this grant application is to generate and test transgenic mouse lines containing ligand regulated Cre recombinase driven by 3.6 and 2.3 kb fragments of the rat Collal promoter. Currently available information T2 indicates that a tamoxifen regulated Cre, Cre-ER , is probably the most appropriate for our studies, Unmodified Cre recombinase constitutively catalyzes deletion of DNA between loxP sites, however Cre-ER T2 is inactive in the absence of the estrogen analogue tamoxifen. Constructs containing the 3.6 and 2.3 kb Col I al promoter driving Cre-ER T22 will be produced and used to generate transgenic mice. Testing of these transgenic mouse lines will be carried out by crossing them with a ROSA 26 Cre reporter mouse strain in which expression of B-galactosidase (beta-gal) or green fluorescent protein (GFP) is dependent on Cre activity. It is expected that reporter gene expression will be specific to the cell types that express the Collal promoters, and dependent on injection of tamoxifen. We will characterize the minimum dosage and time course of tamoxifen treatment needed to optimally induce the reporter gene. We will also characterize the effects of long-term tamoxifen treatment at this dosage on bone metabolism. The purpose for developing these lines is to enable tissue specific and temporally regulated inactivation of genes that are expressed in osteoblasts and are believed to play important roles in regulating osteoblast function, but whose role in adult animals is not understood. By allowing normal expression of a gene during early development and inactivating the gene in adult osteoblasts, we can learn about the specific role of the gene in adult bone metabolism.

描述（由申请人提供）：本次拨款申请的目标是 生成并测试含有配体调节 Cre 的转基因小鼠品系 由大鼠 Collal 启动子的 3.6 和 2.3 kb 片段驱动的重组酶。 目前可获得的信息 T2 表明他莫昔芬调节 Cre， Cre-ER，可能是最适合我们研究的，Unmodified Cre 重组酶组成型催化 loxP 位点之间 DNA 的删除， 然而，如果没有雌激素类似物，Cre-ER T2 就会失去活性 他莫昔芬。含有 3.6 和 2.3 kb Col I al 启动子的构建体 驱动Cre-ER T22将被生产并用于产生转基因小鼠。 将通过杂交对这些转基因小鼠品系进行测试 ROSA 26 Cre 报告小鼠品系，其中表达 B-半乳糖苷酶 (β-gal) 或绿色荧光蛋白 (GFP) 取决于 Cre 活性。它 预计报告基因表达将针对细胞类型 表达 Collal 启动子，并且依赖于他莫昔芬的注射。我们 将描述他莫昔芬治疗的最小剂量和时程 需要最佳地诱导报告基因。我们还将描述 该剂量的长期他莫昔芬治疗对骨代谢的影响。这 开发这些线的目的是使组织具有特异性和暂时性 成骨细胞中表达的基因的调节失活 据认为在调节成骨细胞功能中发挥重要作用，但其 在成年动物中的作用尚不清楚。通过允许正常表达 早期发育过程中的基因并使成体成骨细胞中的基因失活， 我们可以了解该基因在成人骨代谢中的具体作用。