Cited by 559RNA and protein synthesis mechanismsRNA Interference and Gene DeliveryRNA Research and Splicing

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

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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Crossref · 10.1006/mthe.2000.0050

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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
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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