Study for the Production of Immunodeficiency Animal for Xenotransplantation

Transgenes in HSY-TK gene driven by the lck promoter was tested for the expression in immune cells (Jurkat cells) to apply xenotransplantation of human cells into transgenic animals for the potential use of the proliferation or differentiation of human stem cells in the large animal such as an pig. Also, lck-CFP gene was used for transfection experiment into Jurkat cell to confirm the proper regulation of lck promoter for transgene expression in the T cells. Transfection of lck-GFP gene into Jurkat ceils induced CFP expression in transfected cells. The expression of Ick-TK and Ick-CFP genes was confirmed by RT-PCR using RNAs extracted from Jurkat cells, When Jurkat cells transfected with TK and CFP genes were selected against G418 or gancyclovir treatments, Jurkat cells transfected with TK gene were not proliferated in G4i8 and gancyclovir medium while intact cells or cells transfected with CFP gene could grow in gancyclovir medium. However, Jurkat cells transfected with TK or GFP gene were proliferated in G418 medium probably due to Neo$^{r}$ gene in the vector. Gancyclovir treatment destroyed Jurkat cells expressing TK gene indicating that T-cells expressing TK gene can be selectively eliminated by TK gene expression driven by lck promoter.