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.
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