CORE: MICROINJECTION CORE FACILITY

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。转基因/微注射核心设施的目的是为这项建议的研究人员以及OMRF和OUHSC的其他研究人员提供复杂的哺乳动物分子遗传学服务。服务包括通过将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，以及通过将胚胎植入寄养接受者来重新衍生小鼠品系。这些程序中的所有微注射和动物操作都由微注射核心设备执行。候选转基因幼崽(来自受精卵注射)、嵌合体(来自胚泡注射)和重新衍生的幼崽被转移到请求调查者进行分子分析和进一步育种。受精卵注射的应用是表达正常情况下不在特定组织或细胞类型中表达的基因的一代小鼠，表达突变的基因，在小鼠中表达人类基因，或在非整倍体模型中表达额外的基因副本。根据实验参数的不同，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，也可以是携带自身调控元件的大基因组区域，如BAC载体。胚胎干细胞被用来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，以及基因在时间和/或空间表达有条件改变的品系(条件性敲除)。在两种情况下，重新衍生是必要的：第一，当研究人员需要目前仅作为冷冻胚胎分发的小鼠品系时；第二，当研究人员希望从合作实验室进口小鼠，而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时。一种能够提供分子遗传服务的现代转基因/微注射核心设施为这一Cobre提案中的研究人员以及OMRF、OUHSC和整个俄克拉何马州的研究人员增加了重要的力量。

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.