SBIR-STTR Award

The rat is a major experimental animal for biomedical research in the fields of pharmacology, transplantation, cancer, toxicology, physiology, immunology, aging, neurobiology, and drug discovery and development. The goal of this proposal is to provide a technical means to introduce modifications into the rat genome to produce new animal models, genetically tailored for specific purposes. The goal of the Phase I project is to generate embryonic stem (ES) cells from rat embryos and to demonstrate their developmental pluripotence in vitro and in vivo. ES cell lines will be derived from rat blastocysts and characterized in vitro with differentiation assays, enzymatic and antibody markers, and karyotyping. The developmental pluripotence of candidate cell lines will be demonstrated through generation of chimeric rats by transplanting ES cells to host blastocysts. The chimeras will be bred to demonstrate germ line transmission of the ES cell genotype. In phase II, rat genes will be targeted through homologous recombination to genetically alter the ES cells, and these cells will be used to generate germ line chimeras. The chimeras will be bred to produce genetically modified animals.

Thesaurus Terms:
clone cell, embryo /fetus cell culture, embryonic stem cell, method development genotype, tissue mosaicism genetic manipulation, karyotype, laboratory rat

Mouse embryonic stem (ES) cells inaugurated a new era for mouse geneticists, by allowing them to mutate specific genes and also produce mice carrying large genomic transgenes. If it were possible to manipulate the rat genome in a similar manner through rat ES cells, the powerful database of rat physiology and behavior could be exploited to produce transgenic models of human disease. The Phase I research project demonstrated the feasibility of deriving ES cell lines from rats through development of methods for obtaining sources of ES cell precursors, tissue culture procedures, and in vitro and in vivo methods to assess the developmental potential of cell lines. For the Phase II project, these methods will be applied to further assess the two cell lines derived in the Phase I project and to rapidly analyze new candidate lines. The goals of the Phase II project are to produce a germ-line competent rat ES cell line; to develop transfection techniques for the cells; and to use the cells to begin production of a At model for Alzheimer's disease, with a new YAC transgenesis method developed at GenPharm.National Institute of Child Health and Human Development (NICHD)