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


CONCLUSIONS
rAAV-EPO expressed biologically active hEpo effectively, suggesting that it is a suitable gene expression tool for further study in vivo experiments.
Diao
Yong
Y
Institute of Xinzhong Drug Research, China Pharmaceutical University, Nanjing 210009, China.
Xu
Rui-An
RA
Wang
Guang-Ji
GJ
Zhao
Xiao-Chen
XC
D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D004622:Embryo, Mammalian; D004921:Erythropoietin / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins
D018076:DNA, Complementary; D004274:DNA, Recombinant; D012333:RNA, Messenger; D011994:Recombinant Proteins; D004921:Erythropoietin