Core--Animal and Molecular Biology Facility

动物和分子生物学核心的目标是通过1)保持和培育转基因和基因敲除(KO)小鼠群体及其基因分型，2)冷冻保存这些小鼠品系的胚胎和精子，以及3)进行原位杂交检测用于基因表达研究的细胞特异性mRNA定位，为参与PPG的研究人员提供服务。参与调查的人员已经以自己的身份拥有了30多个转基因和KO小鼠品系。其中许多已经被证明是癌症研究的有用模型。因此，这个核心的初始目标将是集中和维护这些线路。然而，老鼠群体的维持需要持续的监督。 而且这是一项劳动密集型的工作。这一核心的建立将减少个别调查员的工作量，而且将具有极高的成本效益。然而，在最初建立之后，为了该计划的目的，将添加更多行。因此，将畜牧业集中在这个核心上，并有专门的人员履行日常职责，将提高这个项目的质量和效率。这个核心的另一个重要作用是通过冷冻保存胚胎和精子来保存这些转基因/KO品系，以避免任何无意中的品系丢失，供科学家未来使用。此外，该PPG下的所有项目还涉及与组织/细胞中基因表达的细胞特异性信使核糖核酸分析有关的研究。在过去的12年里，我们一直在参与开发和调整一种非常 灵敏的原位杂交技术，用于研究几乎所有类型的冰冻组织切片的基因表达。使用这项技术，我们已经能够检测到由异种细胞类型组成的组织上极低的mRNAs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)对转基因小鼠进行基因分型和数据库管理；3)进行转基因小鼠品系的杂交育种，包括创建双基因或多基因敲除小鼠；4)为研究人员提供KO小鼠或组织；5)进行KO小鼠胚胎和精子的冷冻保存；6)开发和/或维护各种用作分子的cDNA。 杂交研究中的探针；7)进行原位杂交以检测mRNAs的细胞特异性定位。

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.