Transgenic Core

转基因技术对于单个实验室来说昂贵得令人望而却步，而且需要非常熟练的技术 以及敬业的人员和机构支持。这是因为，首先，许多因素影响很大 随机插入的DNA的表达水平，这导致许多转基因在转录上是沉默的 染色体位置。此外，靶向胚胎干细胞是否会 为生殖系做出贡献。因此，没有转基因设施可以保证转基因的表达，也没有转基因设施可以保证转基因的表达。 靶向的ES细胞将进入生殖系。其次，许多研究人员的研究需要使用 转基因动物并没有在他们那一代中接受过训练。这在一定程度上是这个计划项目的情况 研究人员和他们的实验室人员的专业知识从丰富的经验到很少的经验不等。 经验。因此，UMKC转基因核心中可用的专业知识也得到了 包括在此计划项目中。主任和联席主任将协助首席调查员 设计建筑，教育人员和学生，监督核心的总体运行。这个 这一核心的服务将不仅是产生转基因小鼠，而且将杂交小鼠产生 正确的基因型，并保持小鼠直到准备好接受机械菌株核心或被送到 调查员。这一核心的目的是为本组织成员提供高效和经济的服务 计划项目加上所需的教育部分。核心还将提供以下内容：1) 用于筛选Dmp1和Mepe启动子片段表达的转基因胚胎的产生 骨细胞的选择性和对机械负荷的反应；2)表达8的转基因小鼠的产生 和/或10kb Dmp1-Cre，完整的MEPE或MEPE启动子片段，3)。野生型Cx43和Cx43的产生 截短Cx43小鼠及其在骨细胞中的靶向表达4)Dmp1-loxP和E-11-loxP的产生 5)用不同启动子调控的骨细胞缺失Dmp1或E11的牙线状小鼠 Cre表达；以及6)维持本项目所需的所有Cre小鼠，包括骨钙素-Cre，8 (和/或)kb Dmp1 Cre和3.2 kb Col 1a1 ERT2-Cre小鼠。此外，核心将作为一种资源 为主要调查人员、他们的学生和人员提供关于基因分型和 转基因和基因敲除小鼠的筛选。

Transgenic technologies are prohibitively expensive for individual laboratories and require extremely skilled and dedicated personnel and institutional support. This is because first, many factors greatly affect expression levels of randomly inserted DNA, which leads to transgenes being transcriptionally silent at many chromosomal locations. In addition, there is a considerable risk as to whether targeted ES cells will contribute to the germline. As a result, no transgenic facility will guarantee transgene expression, nor that targeted ES cells will go to germline. Secondly, many investigators whose studies require the use of transgenic animals are not trained in their generation. This is partly the case for this program project where the expertise of investigators and their lab personnel range from extensive experience to very little experience. Therefore, the expertise that is available in the transgenic core at UMKC has also been included in this program project. The Director and Co-Director will assist the principal investigators in the design of constructs, educate personnel and students, and oversee the general operation of the core. The services of this core will not only be to generate transgenic mice, but to cross breed mice to generate the correct genotype, and to maintain the mice until ready for the Mechanical Strain Core or to be sent to the investigator. The purpose of this core is to provide efficient and economical service to the members of the Program Project plus the needed educational component. The core will also provide the following: 1) generation of transgenic embryos for screening of expression of Dmp1 and Mepe promoter fragments for osteocyte selectivity and response to mechanical load; 2) generation of transgenic mice expressing the 8 and/or 10 kb Dmp1-Cre, intact MEPE or MEPE promoter fragments, 3). generation of wild type Cx43 and truncated Cx43 mice and their targeted expression in osteocytes; 4) generation of Dmp1-loxP and E-11-loxP mice; 5) generation of floxed mice lacking Dmp1 or E11 in osteocytes using various promoters regulating Cre expression; and 6) maintainance of all Cre mice needed for this project, including osteocalcin-Cre, 8 (and/or 10) kb Dmp1 Cre, and 3.2 kb Col 1a1 ERt2-Cre mice. In addition, the Core will serve as a resource for consultation and training for prinicipal investigators, their students and personnel for genotyping and screening of transgenic and knockout mice.