Core--Animal and Molecular Biology Facility

核心--动物与分子生物学设施

• 批准号: 6997745
• 负责人: SANJOY K. DAS
• 金额: $ 19.33万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-22 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6997745
• 关键词: animal breeding; biomedical facility; cancer prevention; chemoprevention; complementary DNA; cryopreservation; data management; eicosanoids; embryo /fetus; genetically modified animals; in situ hybridization; laboratory mouse; molecular biology information system; nonsteroidal antiinflammatory agent; prostaglandin endoperoxide synthase; sperm; tissue resource /registry

The objective of the Animal and Molecular Biology core is to serve the participating investigators of this PPG by 1) maintenance and breeding of transgenic and knockout (KO) mouse colonies and their genotyping, 2) cryopreservation of embryos and sperms from these mouse lines, and 3) performing in situ hybridization for detection of cell-specific mRNA localization for gene expression studies. The participating investigators already possess more than 30 lines of transgenic and KO mice on their own capacity. Many of these lines have already shown to be useful as models for cancer research. Therefore, the initial goal of this core will be to centralize and maintain these lines. However, maintenance of mouse colonies needs constant supervision and it is a labor-intensive endeavor. The establishment of this core will reduce the workloads of individual investigators and will be extremely cost-effective. Following the initial establishment, however, additional lines will be added for the purpose of the program. Thus, centralizing animal husbandry in this core and having dedicated personnel to perform routine duties will improve the quality and efficiency of this program. Another important role of this core will be to preserve these transgenic/KO lines by cryopreservation of embryos and sperms simply to avoid any inadvertent loss of lines and for future use by scientists. Additionally, all projects under this PPG also involve studies in relation to the cell-specific mRNA analysis for gene expression in tissues/cells. For the last 12 years, we have been involved to develop and adapt a very sensitive technique of in situ hybridization for gene expression studies for virtually all kinds of frozen tissue sections. Using this technique, we have been able to detect extremely low expression of mRNAs on tissues that are comprised of heterogeneous cell types. Our primary goals are to: 1) perform routine husbandry of transgenic and KO mice used in this program; 2) carry out genotyping of transgenic mice and database management; 3) perform cross-breeding of genetically defined mouse strains including creation of double or multiple gene knock-out mice; 4) provide KO mice or tissues to investigators; 5) perform cryopreservation of embryos and sperms from KO mice; 6) develop and/or maintain various cDNAs for their use as molecular probes in hybridization studies; 7) Perform in situ hybridization to detect cell-specific localization of mRNAs.

动物和分子生物学核心的目的是通过1）通过1）为参与的研究者提供了转基因和基因敲除（KO）小鼠菌落的维持和繁殖，及其基因分型，2）从这些小鼠线中对胚胎和精子的冷冻保存，以及3）进行现场杂交以检测细胞杂交以检测细胞杂种型的基因研究，以进行基因定位。参与的调查人员已经拥有30行的转基因和KO小鼠以其自身的能力。这些线中的许多已经证明是癌症研究模型有用的。因此，该核心的最初目标是集中和维护这些线路。但是，鼠标菌落的维护需要不断监督 这是一项劳动密集型的努力。建立该核心将减少个别调查人员的工作量，并具有极高的成本效益。但是，在初始建立之后，将出于程序目的添加其他行。因此，将畜牧业集中在此核心中，并有专门的人员来执行常规职责，将提高该计划的质量和效率。该核心的另一个重要作用将是通过冷冻保存胚胎和精子来保留这些转基因/KO线，只是为了避免任何无意的线路损失以及科学家将来使用。此外，该PPG下的所有项目还涉及与细胞特异性mRNA分析有关组织/细胞中基因表达的研究。在过去的12年中，我们参与了发展和适应非常 对于几乎所有类型的冷冻组织切片的基因表达研究，原位杂交的敏感技术。使用这种技术，我们能够检测到由异质细胞类型组成的组织上mRNA的极低表达。我们的主要目标是：1）执行该程序中使用的转基因和KO小鼠的常规饲养； 2）进行转基因小鼠和数据库管理的基因分型； 3）对遗传定义的小鼠菌株进行杂交，包括产生双重或多个基因敲除小鼠； 4）向研究人员提供KO小鼠或组织； 5）对KO小鼠的胚胎和精子进行冷冻保存； 6）开发和/或维持各种cDNA作为分子使用 杂交研究的探针； 7）进行原位杂交以检测mRNA的细胞特异性定位。