Production of human CD59 cDNA transgenic founder mice——an animal model for xenotransplantation

Objective The founder transgenic mice were generated to build a line of transgenic mice expressing human CD59 protein as an animal model for further study of xenograft rejection. Methods The transgene pEGFP-C1-OMT-CD59 containing human CD59 cDNA and green fluorescent protein gene (GFP) was constructed. The transgene was purified and collected as 3.3 kb gene fragments, before being microinjected into fertilized eggs. The newborn F 0 founder transgenic mice with positive CD59 cDNA were first screened by using polymerase chain reaction (PCR) to examine the DNA in their tail tissue, and then further identified by Southern hybridization with CD59 cDNA as the probe. Results Totally 590 fertilized eggs were microinjected and transferred into 24 recipient mice, 73 mice were born (with an embryo survival rate of 12.4%). PCR showed 8 of the 73 (6♂∶2♀) were positive, while Southern hybridization showed 2 of the 8 (1♂∶1♀) were positive, with the copy numbers of CD59 cDNA integrated being 3 and 10. Conclusion The human CD59 cDNA transgenic founder mice were successfully generated.