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IRES-Dependent Second Gene Expression Is Significantly Lower Than Cap-Dependent First Gene Expression in a Bicistronic Vector

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Mizuguchi, Hiroyuki, Xu, Zhili, Ishii-Watabe, Akiko, Uchida, Eriko, Hayakawa, Takao·2000·Molecular Therapy

DOI 10.1006/mthe.2000.0050

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Abstract

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
ORCID: Mizuguchi, Hiroyuki
Xu, Zhili
Hayakawa, Takao
Venue: Molecular Therapy
Publisher: Elsevier BV
Published: 2000-04
Volume: 1
Issue: 4
Pages: 376-382
ISSN: 1525-0016 (ISSN-L)
DOI: 10.1006/mthe.2000.0050
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: cc-by-nc-nd
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
creativecommons.org/licenses/by-nc-nd/4.0/ (vor, 6033 d)
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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IRES-Dependent Second Gene Expression Is Significantly Lower Than Cap-Dependent First Gene Expression in a Bicistronic Vector

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