Construction of new amplifier expression vectors for high levels of IL-2 gene expression.

T. C. Tsang, J. L. Brailey, F. H. Vasanwala, R. S. Wu, F. Liu, P. R. Clark, L. Meade-Tollin, L. Luznick, A. T. Stopeck, E. T. Akporiaye, D. T. Harris

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

The success of IL-2 gene therapy in cancer is in part dependent on the development of high level IL-2 gene expression vectors. Currently, expression vectors based on the human cytomegalovirus (CMV) promoter give the highest levels of expression. We have attempted to construct new IL-2 expression vectors to test whether gene expression can be further increased. The first approach was to use the new SR-alpha promoter to control IL-2 gene expression. The second approach was to combine the Tat transcription activator gene and the HIV 1 and 2 promoters in the same construct so that the levels of gene expression can be amplified. Transient transfection results using the human colon cancer cell line SW480 showed that the SR-alpha promoter yields similar levels of activity as the CMV promoter. However, the HIV 1 and 2 promoter-based amplifier constructs produced 11 and 28 times more secreted IL-2 than the CMV promoter control. The augmented activity of the amplifier constructs was dependent on the presence of the Tat gene and the transcriptional units must be placed in the same orientation. Reducing the size of the vectors by elimination of the neomycin selectable marker did not increase the activity of the constructs.

Original languageEnglish (US)
Pages (from-to)295-300
Number of pages6
JournalInternational journal of molecular medicine
Volume5
Issue number3
DOIs
StatePublished - Mar 2000

ASJC Scopus subject areas

  • Genetics

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    Tsang, T. C., Brailey, J. L., Vasanwala, F. H., Wu, R. S., Liu, F., Clark, P. R., Meade-Tollin, L., Luznick, L., Stopeck, A. T., Akporiaye, E. T., & Harris, D. T. (2000). Construction of new amplifier expression vectors for high levels of IL-2 gene expression. International journal of molecular medicine, 5(3), 295-300. https://doi.org/10.3892/ijmm.5.3.295