Abstract
The use of DNA vectors to elicit an immune response has produced a lot of interest. Unfortunately, one of the limiting factors has been the problem of gene expression. In order to obtain a strong expression of the vaccinating gene, several steps are necessary. The vector has to be delivered in such a way that it is not being degraded by the immune nor by the hepatic system; it has also to enter efficiently the targeted cells; and it must be expressed in the appropriate compartment of the cells at a high level. For these reasons, we have developed a gene expression vector that contains a T7 autogene and is being expressed in the cytoplasm of the cells (1,2). We will describe this system and two possible applications: infectious disease vaccination and tumor ablation. The latter application may be combined with DNA vaccination against cancer cells.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chen, X., Li, Y., Xiong, K., and Wagner, T. (1994) A self-initiating eukaryotic transient gene expression system based on cotransfection of bacteriophage T7 RNA polymerase and DNA vectors containing a T7 autogene. Nucleic Acids Res. 22, 2114ā2120.
Chen, X., Li, Y., Xiong, K., Xie, Y., Aizicovici, S., Snodgrass, R., et al. (1995). A novel nonviral cytoplasmic gene expression system and its implications in cancer gene therapy. Cancer Gene Ther. 2, 281ā289.
Dunn, J. J., Krippl, B., Bernstein, K. E., Westphal, H., and Studier, F. W. (1988) Targeting bacteriophage T7 RNA polymerase to the mammalian cell nucleus. Gene 68, 259ā266.
Dubendorff, J. W. and Studier, F. W. (1991) Controlling basal expression in an inducible T7 expression system by blocking the target T7 promoter with lac repressor. J. Mol. Biol. 219, 61ā68.
Selby, M. J., Doe, B., and Walker, C. M. (1997) Virus-specific cytotoxic T-lymphocyte activity elicited by coimmunization with human immunodeficiency virus type-1 genes regulated by the bacteriophage T7 promoter and T7 RNA polymerase protein. J. Virol. 71, 7827ā7831.
Chen, X., Li, Y., Xiong, K., Aizicovici, S., Xie, Y., Zhu, Q., et al. (1998) Cancer gene therapy by direct injection of a nonviral T7 vector encoding a thymidine kinase gene. Hum. Gene Ther. 9, 729ā736.
Winter, G., Fields, S., and Brownlee, G. (1981) Nucleotide sequence of the hemagglutinin gene of a human influenza virus H1 subtype. Nature 292, 72ā75.
Caton, A. J., Brownlee, G. G., Yewdell, J. W., and Gerhard, W. (1982) The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin (H1 subtype). Cell 31, 417ā427.
Kunkel, T. A. (1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc. Natl. Acad. Sci. USA 82, 488ā492.
Sanger, F., Nicklen, S., and Coulson, A. R. (1977) DNA sequencing with chainterminating inhibitors. Proc. Natl. Acad. Sci. USA 74, 5463ā5467.
Doe, B., Steimer, K. S., and Walker, C. M. (1994) Induction of HIV-1 envelope (gp120)-specific cytotoxic T lymphocyte responses in mice by recombinant CHO cell-derived gp120 is enhanced by enzymatic removal of N-linked glycan. Eur. J. Immunol. 24, 2369ā2376.
Studier, F. W. (1991) Use of bacteriophage T7 lysozyme to improve an inducible T7 expression system. J. Mol. Biol. 219, 37ā44.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
Ā© 2000 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Sturtz, F.G., Li, Y., Shulok, J., Snodgrass, H.R., Chen, Xz. (2000). A Nonviral Cytoplasmic T7 Autogene System and Its Applications in DNA Vaccination. In: Lowrie, D.B., Whalen, R.G. (eds) DNA Vaccines. Methods in Molecular Medicineā¢, vol 29. Humana Press. https://doi.org/10.1385/1-59259-688-6:323
Download citation
DOI: https://doi.org/10.1385/1-59259-688-6:323
Publisher Name: Humana Press
Print ISBN: 978-0-89603-580-5
Online ISBN: 978-1-59259-688-1
eBook Packages: Springer Protocols