High-level expression of exogenous genes by replication-competent retrovirus vectors with an internal ribosomal entry site

Gene
M MurakamiH Iba

Abstract

We report the construction of two types of Rous sarcoma virus (RSV)-based replication-competent avian retrovirus vectors, IR1 and IR2 to express an exogenous gene at a very high level. In these vectors, the internal ribosomal entry site (IRES) derived from encephalomyocarditis virus (EMCV) was inserted between the env gene and an exogenous gene. The IR1 vector retains the splicing acceptor site that is present in the downstream of the env gene while the IR2 vector lacks it. Using a v-fos mutant (v-fos-CD3) as an example of exogenous genes, we show here that both IR1 and IR2 vectors expressed the gene product, CD3, at expression levels 5- and 8-fold higher than that of their parental vector without IRES, respectively. These vectors were moderately stable and kept a high-level expression of CD3 for at least three passages through the cells. Analysis of viral transcripts indicate that exogenous genes carried by both IR vectors were translated exclusively from the IRES that is present in all the species of the viral transcripts. High-level expression of exogenous genes was also observed in the case of the Hoxa-13 gene in the IR1 vector or the fra-2 gene in the IR2 vector, indicating that the extremely high-level expression characte...Continue Reading

References

Jan 31, 1991·Biochemical and Biophysical Research Communications·T YoshidaH Iba
May 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·H NishinaH Iba
Jul 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·H IbaH Hanafusa
Jun 11, 1993·Nucleic Acids Research·T YoshidaH Iba
Apr 15, 1997·Proceedings of the National Academy of Sciences of the United States of America·H WatanabeH Iba

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Citations

May 13, 1999·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·T KamedaH Iba
Jul 26, 2000·Development, Growth & Differentiation·H Iba
Dec 10, 2009·The Journal of Experimental Medicine·Shintaro KamizonoTak W Mak
May 10, 2006·Development, Growth & Differentiation·Dwi Listyorini, Sadao Yasugi
Mar 23, 2005·Fukushima Journal of Medical Science·Noboru Sato
Nov 10, 2000·Biochemical and Biophysical Research Communications·M UiH Iba
Aug 14, 2003·Mechanisms of Development·Kimiko FukudaSadao Yasugi
May 29, 2003·The Journal of Biological Chemistry·Mitsuko SuzukiAtsushi Kuroiwa

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