Fusagene vectors: A novel strategy for the expression of multiple genes from a single cistron

J. Gäken, J. Jiang, K. Daniel, E. Van Berkel, C. Hughes, M. Kuiper, D. Darling, M. Tavassoli, J. Galea-Lauri, K. Ford, M. Kemeny, S. Russell, F. Farzaneh

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Transduction of cells with multiple genes, allowing their stable and co-ordinated expression, is difficult with the available methodologies. A method has been developed for expression of multiple gene products, as fusion proteins, from a single cistron. The encoded proteins are post-synthetically cleaved and processed into each of their constituent proteins as individual, biologically active factors. Specifically, linkers encoding cleavage sites for the Golgi expressed endoprotease, furin, have been incorporated between in-frame cDNA sequences encoding different secreted or membrane bound proteins. With this strategy we have developed expression vectors encoding multiple proteins (IL-2 and B7.1, IL-4 and B7.1, IL-4 and IL-2, IL-12 p40 and p35, and IL-12 p40, p35 and IL-2). Transduction and analysis of over 100 individual clones, derived from murine and human tumour cell lines, demonstrate the efficient expression and biological activity of each of the encoded proteins. Fusagene vectors enable the co-ordinated expression of multiple gene products from a single, monocistronic, expression cassette.

Original languageEnglish (US)
Pages (from-to)1979-1985
Number of pages7
JournalGene Therapy
Volume7
Issue number23
DOIs
StatePublished - 2000

Keywords

  • Furin
  • Fusion protein
  • Protein expression
  • Retroviral vectors

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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    Gäken, J., Jiang, J., Daniel, K., Van Berkel, E., Hughes, C., Kuiper, M., Darling, D., Tavassoli, M., Galea-Lauri, J., Ford, K., Kemeny, M., Russell, S., & Farzaneh, F. (2000). Fusagene vectors: A novel strategy for the expression of multiple genes from a single cistron. Gene Therapy, 7(23), 1979-1985. https://doi.org/10.1038/sj.gt.3301341