Adeno-associated virus mediated expression of human erythropoietin in vitro.

ABSTRACT

AIM: To investigate the expression of human erythropoietin (hEpo) in 293 cell line using adeno-associated virus (AAV) vector, and provide information for its potential application in human gene therapy. METHODS: The human Epo cDNA was inserted into the AAV cassettes, and recombinant adeno-associated virus (rAAV-EPO) vectors were generated by cotransfection methods without helper virus. The expression of hEpo was detected by RT-PCR and ELISA methods. RESULTS: A recombinant adeno-associated virus expressing hEpo was constructed and generated successfully. Positive signal of mRNA was detected from 293 cells transduced with rAAV-EPO, and the hEpo level in supernatant reached 500 U/L. CONCLUSION: rAAV-EPO expressed biologically active hEpo effectively, suggesting that it is a suitable gene expression tool for further study in vivo experiments.