HIGHLY EFFICIENT GENE TARGETING VECTOR AND GENE TARGETING METHOD TOWARD EPITHELIAL CELL LINE

摘要

In the course of constructing a ZO-1 gene knockout mouse, a gene targeting vector causing homologous recombination at a high possibility of 90% or more is developed. As the results of studies on the optimum conditions for electroporation of epithelial cells with the use of this targeting vector, the optimum electroporation conditions enabling efficient gene targeting toward mouse EpH4, which is an epithelial cell line, are successfully determined. Use of this vector makes it possible to easily introduce a foreign gene to the Z0-1 allele of an ES cell. Since it is unnecessary to take the effects of genome structure into consideration, this method is expected as contributing to the solution of problems occurring in the existing methods of constructing transgenic mice and preparing transformants with stable cells.