Retroviral Vectors Pseudotyped with Chimeric Vesicular Stomatitis Virus Glycoprotein for Antibody-Dependent Gene Transduction

Abstract

Retroviral vectors are a powerful tool for stable gene transfer in animal cells because of their ability to integrate the target gene into the host genome. Above all, retroviral vectors pseudotyped with vesicular stomatitis virus glycoprotein G (VSV-G) have been used widely because of their broad host range specificity. These vectors can infect various cells regardless of species (pantropic), since the receptors for VSV-G are phospholipids which are observed ubiquitously in membrane lipids. In addition, pseudotyping with VSV-G improves the mechanical strength of viral vectors, and viral titers can be increased by ultracentrifugation. However, the in vivo use of pantropic viral vectors is limited due to the low transduction efficiency of target cells and possible side-effects. In the present study, an expression plasmid for chimeric VSV-G, consisting of a ZZ fragment derived from Staphylococcal protein A fused to the N-terminus of VSV-G (ZZ-VSV-G), was constructed to produce viral vectors capable of antibody-dependent gene transduction. Gammaretroviral and lentiviral vectors pseudotyped with ZZ-VSV-G were produced without the loss of antibody-binding activity. The production of infectious viral particles was promoted by the addition of an expression plasmid for native VSV-G and antibody-dependent gene transduction was achieved using plates coated with antibodies. This system may be useful for genetic transduction of cells expressing specific proteins on their surface, and for screening of antibodies specific for cell surface receptors.