Adenovirus-mediated transfer of the amphotropic retrovirus receptor cDNA increases retroviral transduction in cultured cells.
The presence of functional amphotropic receptors on the cell surface is necessary for amphotropic retrovirus-mediated gene transfer. A recombinant adenoviral vector that expresses the receptor for amphotropic retrovirus (RAM) was constructed and used to express the receptor cDNA in different cell types in culture. Transfer of the RAM cDNA increased amphotropic retroviral-mediated transfer from 0 to 60% in Chinese hamster ovary cells. RAM expression increased retroviral transduction four- to eight-fold from 2-4% to 18%-35% in HeLa, Namalva, and X63 cells, but had no effect on 208F and HepG2 cells which have high baseline retroviral transduction rates of about 50%. For the purpose of application to ex vivo gene therapy, primary mouse hepatocytes were studied in a similar manner. Hepatocytes had a baseline transduction efficiency of about 40% and did not have increased rates of retroviral-mediated gene transfer with expression of recombinant RAM. This recombinant adenoviral vector conferred infection of amphotropic retrovirus into cells that were relatively resistant to infection, thus offering a rapid and easy method to stably introduce genes into these cell lines.
Lieber
A
A
Markey Molecular Medicine Center, Department of Medicine, University of Washington, Seattle 98195, USA.
Vrancken Peeters
M J
MJ
Kay
M A
MA
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008099:Liver / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D028061:Phosphate Transport Proteins; D011991:Receptors, Virus / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics*; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters; D014161:Transduction, Genetic
D018076:DNA, Complementary; D028061:Phosphate Transport Proteins; D011991:Receptors, Virus; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters