CORE: MICROINJECTION CORE FACILITY

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。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。所列机构为中心，不一定是研究者所在机构。转基因/显微注射核心设施的目的是为本提案的研究人员以及OMRF和OUHSC的其他研究人员提供先进的哺乳动物分子遗传服务。服务包括通过将DNA引入受精卵以及通过将基因操作的胚胎干细胞注射到胚泡中来生产基因工程小鼠，以及通过将胚胎植入寄养受体来重新衍生小鼠品系。这些程序中的所有显微注射和动物操作均由显微注射核心设施进行。将候选转基因幼仔（来自受精卵注射）、嵌合体（来自胚泡注射）和再衍生幼仔转移给提出请求的研究者进行分子分析和进一步育种。受精卵注射的应用是产生表达通常在特定组织或细胞类型中不表达的基因、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。根据实验参数，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形式，或者是在载体如BAC中携带其自身调控元件的大基因组区域。胚胎干细胞用于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，以及具有条件性改变的基因的时间和/或空间表达（条件性敲除）的品系。在两种情况下，重新衍生是必要的：第一，当研究人员需要一个目前只以冷冻胚胎形式分发的小鼠品系时，第二，当研究人员想要从合作实验室进口小鼠时，小鼠携带已知的病原体，需要通过重新衍生去除这些病原体。一个现代化的转基因/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，调查人员在这个COBRE的建议，以及调查人员在OMRF，OUHSC，并在整个俄克拉荷马州。

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 Transgenic/Microinjection Core Facility is to provide sophisticated mammalian molecular genetic services to the investigators of this proposal, as well as other investigators at OMRF and OUHSC. Services 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, and the rederivation of mouse strains through implantation of embryos into foster recipients. All microinjections and animal manipulations in these procedures are performed by the Microinjection Core Facility. Candidate transgenic pups (from zygote injections), chimeras (from blastocyst injections), and rederived pups are transferred to the requesting investigator for molecular analysis and further breeding. Applications for zygote injection are the generation of mice expressing a gene which normally is not 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. Depending on experimantal parameters, injected DNA can be in the form of 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. Embryonic stem cells are used for 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, and strains with conditionally altered temporal and/or spatial expression of genes (conditional knock-out). Rederivation is necessary in two situations: First, when investigators need a mouse strain which currently is only distributed as frozen embryos, and second, when investigators want to import mice from collaborating laboratories and the mice carry known pathogens, necessitating removal of these pathogens through rederivation. A modern Transgenic/Microinjection Core Facility able to provide molecular genetic services adds important strengths to investigators within this COBRE proposal as well as to investigators at OMRF, OUHSC, and throughout the state of Oklahoma.