CORE: MICROINJECTION CORE FACILITY

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 显微注射核心设施的目的是提供先进的哺乳动物分子遗传服务。 这些包括通过将DNA引入受精卵以及通过将遗传操作的胚胎干细胞注射到胚泡中来生产基因工程小鼠（1），以及通过将胚胎植入寄养受体来重新衍生小鼠品系（2）。将DNA注射到受精卵中的应用实例包括产生表达在特定组织或细胞类型中通常不表达的基因、表达突变基因、在小鼠中表达人基因或表达基因的额外拷贝（例如在非整倍体模型中）的小鼠（3，4）。根据实验的设计，待注射的DNA可以是携带受特定启动子控制的基因编码区的质粒，或在载体如BAC、P1克隆或YAC中携带其自身调控元件的大基因组区域（5）。 需要胚胎干细胞途径的应用是产生携带目的基因的无效等位基因（敲除品系）、基因的突变等位基因（敲入品系）、确定的染色体缺失、复制或易位的小鼠品系，或具有条件性改变的基因的时间和/或空间表达的品系（条件性敲除）（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以冷冻胚胎形式分布的小鼠品系时，重新衍生是必要的。这种情况越来越严重，因为国家小鼠突变资源，如在杰克逊实验室，不能保持在研究界产生的所有基因工程菌株的育种种群。第二种需要重新推导的情况发生在研究人员想要从合作实验室进口小鼠并且小鼠携带已知病原体时。 在这种情况下，病原体通过再衍生被去除。随着研究实验室交换材料的频率越来越高，通过再衍生进行纯化的需求也在增加。 1998年，俄克拉荷马州医学研究基金会（OMRF）在俄克拉荷马州建立了一个用于生产基因工程小鼠的显微注射核心设施。 该设施使整个俄克拉荷马州的生物医学研究人员可以利用老鼠的复杂分子工程。 国内没有其他类似的设施。 一个现代化的小鼠显微注射核心设施能够提供分子遗传服务，增加了重要的优势，在OMRF的研究人员，在OUHSC，并在整个国家的研究人员。 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研究的宝贵研究工具，为定义疾病的病理生物学和在发育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).