CORE: MICROINJECTION CORE FACILITY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 微注射核心设施的目的是提供复杂的哺乳动物分子遗传服务。这些措施包括通过将DNA导入受精卵以及通过将经过基因操作的胚胎干细胞注入囊胚来生产基因工程小鼠(1)，以及通过将胚胎植入寄养接受者来重新衍生小鼠品系(2)。将DNA注入受精卵的应用例子包括：产生表达特定组织或细胞类型中不正常表达的基因的小鼠，表达突变的基因，在小鼠中表达人类基因，或表达额外的基因副本，如在非整倍体模型中(3，4)。根据实验的设计，要注射的DNA可以是携带特定启动子控制下的基因编码区的质粒，也可以是携带其自身调控元件的大基因组区域，如BAC、P1克隆或YAC(5)。需要胚胎干细胞路线的应用是创造携带感兴趣基因的零等位基因的小鼠品系(敲除品系)、基因突变的等位基因(敲入品系)、明确的染色体缺失、复制或易位，或具有有条件改变的基因的时间和/或空间表达的品系(条件性剔除)(6-10)。在两种情况下需要重新派生。首先，当研究人员需要一种目前仅作为冷冻胚胎分布的小鼠品系时，重新衍生是必要的。这种情况越来越严重，因为国家老鼠突变资源，如杰克逊实验室，无法维持研究界生产的所有基因工程菌株的繁育库存。第二个需要重新衍生的案例发生在调查人员希望从合作实验室进口老鼠，而这些老鼠携带已知病原体时。在这种情况下，病原体通过重新派生来去除。随着研究实验室交换材料的频率越来越高，通过再衍生进行提纯的需求也在增加。 1998年，俄克拉荷马州医学研究基金会(OMRF)建立了一个显微注射核心设施，用于在俄克拉荷马州生产基因工程小鼠。该设施为俄克拉荷马州的生物医学研究人员提供了复杂的小鼠分子工程。在该州内没有其他类似的设施。 能够提供分子遗传服务的现代小鼠微注射核心设备为OMRF、OUHSC和全州的研究人员增加了重要的力量。我们相信，这些复杂的哺乳动物分子遗传学将继续成为基础和临床导向研究的宝贵研究工具，为定义疾病的病理生物学和在发展良好的哺乳动物模型中评估治疗效果提供技术(11)。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The purpose of the Microinjection Core Facility is to provide sophisticated mammalian molecular genetic services. These include the production of genetically engineered mice through introduction of DNA into zygotes as well as through injection of genetically manipulated embryonic stem cells into blastocysts (1), and the rederivation of mouse strains through implantation of embryos into foster recipients (2). Examples of applications for injection of DNA into zygotes include the generation of mice expressing a gene not normally expressed in a particular tissue or cell type, expressing mutated genes, expressing human genes in mice, or expressing additional copies of genes such as in aneuploid models (3,4). Depending on the design of the experiments, the DNA to be injected can be a plasmid carrying the coding region of a gene under control of a specific promoter, or large genomic regions carrying their own regulatory elements in vectors such as BACs, P1 clones, or YACs (5). Applications requiring the embryonic stem cell route are the creation of mouse strains carrying a null allele of the gene of interest (knock-out strains), a mutated allele of a gene (knock-in strains), defined chromosomal deletions, duplications, or translocations, or strains with conditionally altered temporal and/or spatial expression of genes (conditional knock-out) (6-10). Rederivation is necessary in two situations. First, rederivation is necessary when investigators need a mouse strain that is currently distributed only as frozen embryos. This is increasingly the case as National Mouse Mutant Resources, such as at the Jackson Laboratory, cannot maintain breeding stocks of all genetically engineered strains produced within the research community. The second case requiring rederivation occurs when investigators want to import mice from collaborating laboratories and the mice carry known pathogens. In this case, pathogens are removed through rederivation. The need for purification through rederivation is also increasing, as research laboratories exchange materials at increasing frequencies. In 1998 the Oklahoma Medical Research Foundation (OMRF) established a Microinjection Core Facility for the production of genetically engineered mice in Oklahoma. This facility makes available the sophisticated molecular engineering of the mouse to biomedical researchers throughout Oklahoma. There exists no other comparable facility within the state. A modern Mouse Microinjection Core Facility able to provide molecular genetic services adds important strengths to investigators at OMRF, at OUHSC, and to investigators throughout the state. We believe these sophisticated mammalian molecular genetics will continue to be an invaluable research tool for both basic and clinically oriented research, providing the technology for defining the pathobiology of diseases and for evaluating therapeutic efficacy in a well-developed mammalian model (11).