IRES-Dependent Second Gene Expression Is Significantly Lower Than Cap-Dependent First Gene Expression in a Bicistronic Vector
Open accessAbstract
The internal ribosome entry site (IRES) has been widely used to coexpress heterologous gene products by a message from a single promoter. However, little is known about the efficiency of IRES-dependent second gene expression in comparison with that of first gene expression. This study was undertaken to characterize the relative expression of IRES-dependent second gene in a bicistronic vector, which was derived from the 5' untranslated regions of the encephalomyocarditis virus (EMCV). IRES-dependent second gene expression was compared with cap-dependent first gene expression in several cultured cell lines and in mouse liver in vivo. The expression of the IRES-dependent second gene ranged from 6 to 100% (in most cases between 20 and 50%) that of the first gene. Second gene expression in a plasmid without the IRES was 0.1-0.8% (with some exceptions) that of the first gene. These findings have important implications for the use of IRES, i.e., care should be taken regarding the decreased capacity of IRES-dependent downstream gene expression as well as in determining which gene should be positioned as the first or second gene in a bicistronic vector.
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- Authors
- Mizuguchi, H., Xu, Z., Ishii-Watabe, A., Uchida, E., & Hayakawa, T.
- Affiliations
- National Institute of Health Sciences (JP)
- Corresponding author
- Mizuguchi, Hiroyuki
- Venue
- Molecular Therapy
- Publisher
- Elsevier BV
- Published
- 2000-04
- Volume
- 1
- Issue
- 4
- Pages
- 376-382
- ISSN
- 1525-0016 (ISSN-L)
- OpenAlex
- W2000853857
- Cited by
- 559
- References
- 34
- Citations by year
- 2026: 12 · 2025: 25 · 2024: 24 · 2023: 37 · 2022: 17 · 2021: 20 · 2020: 23 · 2019: 26 · 2018: 24 · 2017: 15 · 2016: 19 · 2015: 21 · 2014: 24 · 2013: 24 · 2012: 35
- FWCI
- 4.34
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Molecular Therapy (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Keywords
- internal ribosome entry site, bicistronic vector, cap-dependent translation, encephalomyocarditis virus, 5' untranslated region, gene expression efficiency
- MeSH terms
- Alkaline Phosphatase, Animals, Encephalomyocarditis virus, Genes, Genetic Vectors, Cricetinae, HeLa Cells, Humans, L Cells, Liver, Luciferases, Plasmids, RNA Caps, Chloramphenicol O-Acetyltransferase, Gene Expression, CHO Cells, Gene Transfer Techniques, 5' Untranslated Regions, Mice
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