Production of p53 gene knockout rats by homologous recombination in embryonic stem cells

The use of homologous recombination to modify genes in embryonic stem (ES) cells provides a powerful means to elucidate gene function and create disease models1. Application of this technology to engineer genes in rats has previously been impossible in the absence of germline competent ES cells in this species. We have recently established authentic rat ES cells2, 3. Here we report the generation of the first gene knockout rats using the ES cell-based gene targeting technology. We designed a targeting vector to disrupt the tumor suppressor gene p53 (also known as Tp53) in rat ES cells via homologous recombination. p53 gene-targeted rat ES cells can be routinely generated. Furthermore, the p53 gene-targeted mutation in the rat ES cell genome can transmit through the germline via ES cell-rat chimeras to create p53 gene knockout rats. The rat is the most widely used animal model other than humans in biological research4–7. The establishment of gene targeting technology in rat ES cells, in combination with advances in genomics and the vast amount of research data on physiology and pharmacology in this species, now provides a powerful new platform for the study of human disease.