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Adeno-associated virus mediated expression of human erythropoietin in vitro
Yong Diao1, Rui-An Xu, Guang-Ji Wang
1Institute of Xinzhong Drug Research, China Pharmaceutical University, Nanjing 210009, China.
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.