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.
Keywords:
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.