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Use and comparison of different internal ribosomal entry sites (IRES) in tricistronic retroviral vectors.
- Source :
-
BMC biotechnology [BMC Biotechnol] 2004 Jul 27; Vol. 4, pp. 16. Date of Electronic Publication: 2004 Jul 27. - Publication Year :
- 2004
-
Abstract
- Background: Polycistronic retroviral vectors that contain several therapeutic genes linked via internal ribosome entry sites (IRES), provide new and effective tools for the co-expression of exogenous cDNAs in clinical gene therapy protocols. For example, tricistronic retroviral vectors could be used to genetically modify antigen presenting cells, enabling them to express different co-stimulatory molecules known to enhance tumor cell immunogenicity.<br />Results: We have constructed and compared different retroviral vectors containing two co-stimulatory molecules (CD70, CD80) and selectable marker genes linked to different IRES sequences (IRES from EMCV, c-myc, FGF-2 and HTLV-1). The tricistronic recombinant amphotropic viruses containing the IRES from EMCV, FGF-2 or HTLV-1 were equally efficient in inducing the expression of an exogenous gene in the transduced murine or human cells, without displaying any cell type specificity. The simultaneous presence of several IRESes on the same mRNA, however, can induce the differential expression of the various cistrons. Here we show that the IRESes of HTLV-1 and EMCV interfere with the translation induced by other IRESes in mouse melanoma cells. The IRES from FGF-2 did however induce the expression of exogenous cDNA in human melanoma cells without any positive or negative regulation from the other IRESs present within the vectors. Tumor cells that were genetically modified with the tricistronic retroviral vectors, were able to induce an in vivo anti-tumor immune response in murine models.<br />Conclusion: Translation of the exogenous gene is directed by the IRES and its high level of expression not only depends on the type of cell that is transduced but also on the presence of other genetic elements within the vector.
- Subjects :
- Adenocarcinoma genetics
Adenocarcinoma metabolism
Adenocarcinoma pathology
Adenocarcinoma virology
Animals
Antigens, CD genetics
B7-1 Antigen genetics
Bleomycin metabolism
CD27 Ligand
Cell Line, Tumor
Drug Resistance genetics
Gene Expression Regulation genetics
Gentamicins metabolism
Humans
Kidney embryology
Kidney virology
Mammary Neoplasms, Animal genetics
Mammary Neoplasms, Animal metabolism
Mammary Neoplasms, Animal pathology
Mammary Neoplasms, Animal virology
Melanoma genetics
Melanoma metabolism
Melanoma pathology
Melanoma virology
Melanoma, Experimental genetics
Melanoma, Experimental metabolism
Melanoma, Experimental pathology
Melanoma, Experimental virology
Membrane Proteins genetics
Mice
NIH 3T3 Cells chemistry
NIH 3T3 Cells metabolism
RNA, Messenger biosynthesis
Skin Neoplasms genetics
Skin Neoplasms metabolism
Skin Neoplasms pathology
Skin Neoplasms virology
Transduction, Genetic methods
Transgenes genetics
Gene Transfer Techniques
Genes, Viral genetics
Genetic Vectors genetics
Retroviridae genetics
Ribosomes genetics
Viral Structural Proteins genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1472-6750
- Volume :
- 4
- Database :
- MEDLINE
- Journal :
- BMC biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 15279677
- Full Text :
- https://doi.org/10.1186/1472-6750-4-16