Adeno-associated virus mediated expression of human erythropoietin in vitro

Acta Pharmacol Sin. 2002 Jan;23(1):55-8.

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.

MeSH terms

  • Cell Line
  • DNA, Complementary / genetics
  • DNA, Recombinant / genetics
  • Dependovirus / genetics*
  • Dependovirus / metabolism
  • Embryo, Mammalian
  • Erythropoietin / biosynthesis*
  • Erythropoietin / genetics
  • Erythropoietin / metabolism*
  • Genetic Therapy
  • Genetic Vectors / genetics
  • Humans
  • Kidney / cytology*
  • Kidney / metabolism
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Recombinant Proteins

Substances

  • DNA, Complementary
  • DNA, Recombinant
  • RNA, Messenger
  • Recombinant Proteins
  • Erythropoietin