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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 publication

DOI

10.1038/sj.gt.3301341

Type

Journal article

Journal

Gene therapy

Publication Date

12/2000

Volume

7

Pages

1979 - 1985

Addresses

Immune Gene Therapy Programme, Department of Molecular Medicine, The Rayne Institute, London, UK.

Keywords

Lymphocytes, Cells, Cultured, Jurkat Cells, Tumor Cells, Cultured, Animals, Humans, Mice, Retroviridae, Antigens, CD80, Interleukins, Interleukin-2, Interleukin-4, Interleukin-12, Gene Therapy, Transfection, Gene Expression, Genes, Genetic Vectors, Artificial Gene Fusion