Pseudotransduction of hepatocytes by using concentrated pseudotyped vesicular stomatitis virus G glycoprotein (VSV-G)-Moloney murine leukemia virus-derived retrovirus vectors: comparison of VSV-G and amphotropic vectors for hepatic gene transfer
| Autor: | Miao-Liang Liu, Brian Winther, Mark A. Kay |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Genetic enhancement
Immunology Genetic Vectors Microbiology Cell Line Transduction (genetics) Mice Retrovirus Viral Envelope Proteins Virology Murine leukemia virus Gene expression Animals Humans Gene Cells Cultured Membrane Glycoproteins biology Gene Transfer Techniques 3T3 Cells biology.organism_classification beta-Galactosidase Molecular biology Rats Mice Inbred C57BL Liver Cell culture Vesicular stomatitis virus Insect Science Moloney murine leukemia virus Research Article |
| Zdroj: | Journal of virology. 70(4) |
| ISSN: | 0022-538X |
| Popis: | Recombinant retrovirus vectors are widely used for gene transfer studies. The recent development of a pseudotyped Moloney murine leukemia virus vector that contains the G envelope protein from the vesicular stomatitis virus allows for efficient concentration of vector and offers hope for potential use of these vectors for gene expression in vivo. A standard amphotropic vector expressing a serum marker protein, human alpha 1-antitrypsin, was infused into regenerating mouse liver and was 10-fold more efficient at achieving stable gene expression than was an equivalent pseudotyped vector. Discrepant results were obtained with cultured hepatocytes infected with an Escherichia coli beta-galactosidase-producing pseudotype and amphotropic vector. High rates of beta-galactosidase-positive cells were detected with the vesicular stomatitis virus G glycoprotein vector under culture conditions known to be relatively nonpermissive for retrovirus-mediated gene transfer. Subsequent studies demonstrated that beta-galactosidase protein was concentrated and copurified during pseudotype vector preparation, resulting in high rates of protein transfer rather than stable gene transfer, a process referred to as pseudotransduction. The cotransfer of protein with concentrated pseudotyped retroviruses indicates that caution must be used when interpreting gene transduction efficiencies in gene therapy experiments. |
| Databáze: | OpenAIRE |
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