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标是通过 1) 转基因和敲除 (KO) 小鼠群体的维护和育种及其基因分型，2) 冷冻保存这些小鼠品系的胚胎和精子，以及3) 进行原位杂交，检测细胞特异性 mRNA 定位，以进行基因表达研究。参与研究人员已自行拥有30多个转基因和KO小鼠品系。其中许多细胞系已被证明可用作癌症研究模型。因此，这个核心的最初目标将是集中和维护这些线路。然而，小鼠群体的维护需要持续的监督 这是一项劳动密集型工作。该核心的建立将减少个别调查人员的工作量，并且极具成本效益。然而，在最初建立之后，为了该计划的目的，将添加额外的线路。因此，将畜牧业集中到这一核心并由专人履行日常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是通过冷冻胚胎和精子来保存这些转基因/KO 系，以避免任何系的无意丢失并供科学家将来使用。此外，该 PPG 下的所有项目还涉及组织/细胞中基因表达的细胞特异性 mRNA 分析相关的研究。在过去的 12 年里，我们一直致力于开发和调整一个非常 灵敏的原位杂交技术，适用于几乎所有类型的冷冻组织切片的基因表达研究。使用这项技术，我们已经能够检测到由异质细胞类型组成的组织上极低的 mRNA 表达。我们的主要目标是：1）对本项目中使用的转基因和 KO 小鼠进行常规饲养； 2）进行转基因小鼠的基因分型及数据库管理； 3) 对基因确定的小鼠品系进行杂交育种，包括创建双基因或多基因敲除小鼠； 4) 向研究者提供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/或维护各种 cDNA 以用作分子 杂交研究中的探针； 7) 进行原位杂交以检测 mRNA 的细胞特异性定位。