Transgenic Core

转基因技术对于单个实验室来说过于昂贵，并且需要极其熟练的技术人员。 以及专门的人员和机构支持。这是因为，首先，许多因素极大地影响 随机插入DNA的表达水平，这导致转基因在许多情况下转录沉默 染色体位置此外，关于靶向的ES细胞是否会在胚胎干细胞中表达， 对生殖细胞有贡献因此，没有转基因设施能保证转基因表达， 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 转基因动物在它们的世代中没有经过训练。这部分是这个项目的情况， 研究人员和他们的实验室人员的专业知识范围从丰富的经验到很少 体验.因此，UMKC转基因核心中可用的专业知识也被 包括在这个项目中。主任和联席主任将协助主要调查员进行 设计结构，教育人员和学生，并监督核心的一般运作。的 这个核心的服务将不仅是产生转基因小鼠，但杂交小鼠产生的 正确的基因型，并保持小鼠，直到准备好机械应变核心或被送往 调查员这一核心的目的是为会员国提供高效和经济的服务， 计划项目加上所需的教育组成部分。核心还将提供以下内容：1） 产生转基因胚胎，用于筛选Dmp 1和Mepe启动子片段的表达， 骨细胞的选择性和对机械负荷的反应; 2）产生表达8个 和/或10 kb Dmp 1-Cre，完整的MEPE或MEPE启动子片段，3）。产生野生型Cx43和 截短的Cx43小鼠及其在骨细胞中的靶向表达; 4）Dmp 1-loxP和E-11-loxP的产生 小鼠; 5）使用各种启动子调节骨细胞中缺乏Dmp 1或E11的floxed小鼠的产生 Cre表达;和6）该项目所需的所有Cre小鼠的免疫，包括骨钙素-Cre，8 (and/或10 kb Dmp 1 Cre和3.2 kb Col 1a 1 ERt 2-Cre小鼠。此外，核心将作为一种资源， 为主要研究者、他们的学生和基因分型人员提供咨询和培训， 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.