Optimization of Androgen Regulatable, Kidney Proximal Renal Tubule Epithelial Cell Specific Targeted Kap147/HAGT Kpn Hybrid Promoter

Abstract: To study the cell specificity of the Kap147/HAGT Kpn hybrid promoter enhancer combination，the HAGT Kpn fragment was cloned into pGL3 Kap147 vector. Luciferase activity assay after cell transient transfection showed that Kap147/HAGT Kpn hybrid promoter only exhibited activity in OK and LLC ML2 cells that derived from epithelial cells of proximal convoluted tubules, and its activity increased by 7 to 8 fold treated with 20 nmol/L DHT. In order to optimize the hybrid promoter enhancer combination, truncated HAGT Kpn fragments were cloned into pGL3 Kap147 vector. Cell experiments showed that KpnI StuI and MscI KpnI fragments increased expression of Kap147 promoter 1 to 2 fold treated with 20 nmol/L DHT. Kap147/HAGT Kpn hybrid promoter is tissue and cell specific and androgen regulated, while KpnI StuI and MscI KpnI fragments are responsible for the androgen regulatory effect.