The Core will provide support and funding for all animal related activities of Projects 1,2, 3, and Core A. Organizational, supervisory, and budgetary elements will be provided. Experiments in various mouse strains are key for the experiments proposed in Projects 1,2, and 3 to dissect the in vivo roles of TF and components of TF-dependent signaling pathways in vascular biology. Success critically depends on careful and efficient breeding, meticoulous documentation, and genotyping of these knockout, transgenic, and hybrid mouse strains. Backcrossings will be performed by the Core to obtain strains in a defined and stable genetic background. Currently, mouse strains maintained by the Core include TF+/-, TF-/- rescued with a human TF minigene, TF with deleted cytoplasmic domain, PAR1-/-, PAR2-/-, ApoE-/-, LDLR-/-, and TFPI+/-. Efficient PCR based genotyping strategies have been developed for these strains. The Core will be responsible for the maintenance of these and other lines, distribution of animals to the various projects and generating new lines by breeding and genotyping. In addition, mice and rabbits will be provided for Core A to support the generation of specific polyclonal antisera and hybridomas. Most of the mice are housed in the Department of Immunology's vivarium. However, a small-scale breeding program is maintained at the Department of Molecular and Experimental Medicine to safeguard our unique strains against disasters at the Immunology facility, e.g., infectious disease. In addition, frozen embryo stocks of selected mouse lines will be established to further secure these novel and unique resources. The consolidation of all research animal support into a single Core to supervise the conduct of the specific approved protocols of the Program Project has been reviewed and approved by the institutional Animal Research Committee. Activities of the Core will be supervised by Dr. Matthias Riewald. Dr. Riewald has been leading the Animal Core of the current Program Project for more than three years and he is familiar with all aspects of the proposed work. In addition, Dr. Riewald's strong background in molecular biology will assist in the provision of appropriate genotyping techniques for the various mouse lines. Ms. Melissa Szeto will be responsible for maintenance of the mouse lines, record keeping, and genotyping. A lab helper will assist in the day-to-day animal care. A relational database has been designed and created by Dr. Riewald in Microsoft Access for record keeping and rapid analyses of the Program Project's mouse colony. The database facilitates efficient breeding to supply the different Projects with the appropriate numbers of age-and sex-matched animals. Written records of the animal colony are kept as well.
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