Successful transfer of ADA gene in vitro into human peripheral blood CD34+ cells by transfecting EBV-based episomal vectors.
We report a novel non-viral system for transfecting human immature hematopoietic cells in vitro. Epstein-Barr virus (EBV)-based episomal vectors carrying human adenosine deaminase (ADA) gene cDNA were transfected by electroporation into human peripheral blood (PB) CD34+ cells. The transgene-specific mRNA were detected from 37 to 100% of CFU-c (colony forming unit in culture) colonies derived from the transfected cells. A two-fold increase in enzyme activity was also found. These results indicate the successful transfer and expression of genes in human immature hematopoietic cells using the EBV-based episomal vector system.
Satoh
E
E
Department of Microbiology, Kyoto Prefectural University of Medicine, Kamikyo, Japan.
Hirai
H
H
Inaba
T
T
Shimazaki
C
C
Nakagawa
M
M
Imanishi
J
J
Mazda
O
O
D000243:Adenosine Deaminase / Q000096:biosynthesis / Q000235:genetics*; D015703:Antigens, CD / Q000097:blood; D018952:Antigens, CD34 / Q000097:blood*; D002478:Cells, Cultured; D017931:DNA Primers; D018274:Electroporation; D005822:Genetic Vectors; D016179:Granulocyte Colony-Stimulating Factor / Q000627:therapeutic use; D006412:Hematopoietic Stem Cells / Q000201:enzymology* / Q000473:pathology; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D009369:Neoplasms / Q000097:blood / Q000188:drug therapy; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000627:therapeutic use; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D015703:Antigens, CD; D018952:Antigens, CD34; D017931:DNA Primers; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D000243:Adenosine Deaminase