DERIVATION OF RAT EMBRYONIC STEM CELL LINES

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 PROJECT IS TO PROVIDE A TECHNICAL MEANS TO INTRODUCE MODIFICIATIONS INTO THE RATGENOME TO PRODUCE NEW ANIMAL MODELS, GENETICALLY TAILORED FOR SPECIFIC PURPOSES. THE GOAL OF THE PHASE I PROJECT SI TO GENERATE EMBRYONIC STM (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 ANITBODY 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 ALTR THE ES CELLS, AND THESE CELLS WILL BE USED TO GENERATE GERM LINE CHIMERAS. THE CHIMERAS WILLBE BRED TO PRODUCE GENETICALLY MODIFIED ANIMALS.