1. A Tetracycline-regulated Cell Line Produces High-titer Lentiviral Vectors that Specifically Target Dendritic Cells
- Author
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Chupei Zhang, Paul D. Bryson, Pin Wang, and Chi-Lin Lee
- Subjects
General Chemical Engineering ,Immunology ,Genetic Vectors ,CD8-Positive T-Lymphocytes ,Transfection ,General Biochemistry, Genetics and Molecular Biology ,Viral vector ,Cell Line ,Retrovirus ,Virus-like particle ,Humans ,Selectable marker ,Reporter gene ,biology ,General Immunology and Microbiology ,General Neuroscience ,HEK 293 cells ,fungi ,Lentivirus ,Dendritic Cells ,Tetracycline ,biology.organism_classification ,Virology ,HEK293 Cells ,Cell culture ,Plasmids - Abstract
Lentiviral vectors (LVs) are a powerful means of delivering genetic material to many types of cells. Because of safety concerns associated with these HIV-1 derived vectors, producing large quantities of LVs is challenging. In this paper, we report a method for producing high titers of self-inactivating LVs. We retrovirally transduce the tet-off stable producer cell line GPR to generate a cell line, GPRS, which can express all the viral components, including a dendritic cell-specific glycoprotein, SVGmu. Then, we use concatemeric DNA transfection to transfect the LV transfer plasmid encoding a reporter gene GFP in combination with a selectable marker. Several of the resulting clones can produce LV at a titer 10-fold greater than what we achieve with transient transfection. Plus, these viruses efficiently transduce dendritic cells in vitro and generate a strong T cell immune response to our reporter antigen. This method may be a good option for producing strong LV-based vaccines for clinical studies of cancer or infectious diseases.
- Published
- 2013