To define the functions of genes and cell types in vivo, there is no substitute for the power of modifying themouse germline to generate gene knockouts (KO) and mutants or to express new genes transgenically. Foroptimal interpretation, genes need to inactivated conditionally or only in certain tissues, or expressed astransgenes in a tissue specific manner in the native genomic context. Using these techniques, investigatorsworldwide and particularly at Yale, have already been making numerous KO, knockin, transgenic, etc.mouse strains and breeding the modified alleles onto useful genetic backgrounds. These strains, many ofwhich are published, are invaluable resources for research in Rheumatologic diseases, but the maintenanceof them is time consuming and expensive, and one cannot envision the endless collection of more and morestrains of actively breeding mice. Moreover, infection and breeding problems threaten the existence of manystrains, and make the transfer among investigators cumbersome an costly. At the same time, to date thevast majority of engineered mutations have been made by a relatively small number of investigators; thesetechnologies need to be more widely available so that labs that focus on particular genes or processes canhave direct access to modifying them, instead of relying on labs that specialize in making KO mice. Animportant barrier to the technology is in the actual design of KOs and making of constructs as well asconstruction of BAC constructs via homologous recombination. To address these issues, Core B will taketwo approaches. First, the Core will provide cryopreservation (and reconstitution of mice from frozenembryos) capabilities for the cost-effective storage of genetically modified mice. Additionally, the core willinitiate cryopreservation of sperm and the techniques of IVF for inbred mouse lines. This will producesubstantial decreases in costs associated with preservation of mouse strains and will enable distribution bothamong YRDRCC investigators and outside of Yale. Second, to make modern engineering accessible to themajority of YRDRCC member, the core will provide technological expertise in the generation of KO,conditional KO, knock in, BAC and regulated transgenic constructs. The core will also provide strategies forbreeding, genotyping, and initial screening of the resultant mice.
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