TARGETED RESEARCH OF MOUSE Pkdl GENE IN EMBRYONIC STEM CELLS

Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common human genetic disease with an approximate incidence of 1/400 - 1/1000. The disease is characterized by progressive cyst formation and enlargement, typically leading to end stage renal disease (ESRD) by late middle age and accounts for approximately 8-10% of cases of chronic renal failure. About 50% of the patients require renal transplantation or dialysis. Alternatively, ADPKD is associated with cystic growth in other organs, most in the liver.ADPKD is genetically heterogeneous with loci mapped to chromosome l6p I3.3 (PKD1) and to 4q22-23 (PKD2), with the likelihood of a third, unmapped locus (PKD3). PKDI accounts for about 85% of ADPKD and was cloned in 1995. All but 3.5 kb at the 3' end of the transcript is encoded by a region reiterated several times elsewhere on the same chromosome. Mouse pkdl is a single copy on the chromosome 17, and its frill transcript was cloned in 1997.The pathogenesis mechanism of ADPKD is not clear and the treatment of this disease is not efficacy. Animal models of polycystic kidney disease are proving to be extremely useful for studying the molecular basis of renal cyst formation and for finding new drugs to treat this disease. Animal models obtained by chemical induce, spontaneous mutation and eggs microinjection with cancer genes can't satisfy the demand. Transgenic animal models obtained by embryonic stem (ES) cell gene targeting are the best models in which the structure of the gene is modified artificially, and the analogous phenotype of the human genetic disease is obtained.In this study, a pair of primers in the exon 15 of the mouse pkd1 gene was designed with PCGENE software. Genomic DNA extracted from normal mouse was used as a template for PCR amplification. Then a pGEM-pdkl vector was...