The DOI matches its Crossref record; 36 of the 36 references it cites were found in a registry or an index.
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.
Overview
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
Citations
Cites
Efficient selection for high-expression transfectants with a novel eukaryotic vector
Niwa, H., Yamamura, K., & Miyazaki, J. (1991) · Gene · doi:10.1016/0378-1119(91)90434-d · Cited by 5,149
Kozak, M. (1986) · Cell · doi:10.1016/0092-8674(86)90762-2 · Cited by 4,741
The scanning model for translation: an update.
Kozak, M. (1989) · The Journal of Cell Biology · doi:10.1083/jcb.108.2.229 · Cited by 3,485
Improved retroviral vectors for gene transfer and expression.
Miller, A. D., & Rosman, G. J. (1989) · PubMed · Cited by 1,990
Pelletier, J., & Sonenberg, N. (1988) · Nature · doi:10.1038/334320a0 · Cited by 1,953
Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA
Liu, F., Song, Y. K., & Liu, D. (1999) · Gene Therapy · doi:10.1038/sj.gt.3300947 · Cited by 1,728
Jang, S. K., Kräusslich, H., Nicklin, M. J. H., Duke, G. M., Palmenberg, A. C., & Wimmer, E. (1988) · Journal of Virology · doi:10.1128/jvi.62.8.2636-2643.1988 · Cited by 1,390
Effects of Intercistronic Length on the Efficiency of Reinitiation by Eucaryotic Ribosomes
Kozak, M. (1987) · Molecular and Cellular Biology · doi:10.1128/mcb.7.10.3438-3445.1987 · Cited by 537
New mammalian expression vectors
Moss, B., Elroy‐Stein, O., Mizukami, T., Alexander, W. A., & Fuerst, T. R. (1990) · Nature · doi:10.1038/348091a0 · Cited by 523
Emerman, M., & Temin, H. M. (1984) · Cell · doi:10.1016/0092-8674(84)90453-7 · Cited by 434
Jang, S. K., Davies, M. V., Kaufman, R. J., & Wimmer, E. (1989) · Journal of Virology · doi:10.1128/jvi.63.4.1651-1660.1989 · Cited by 413
Elroy‐Stein, O., Fuerst, T. R., & Moss, B. A. (1989) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.86.16.6126 · Cited by 405
Cullen, B. R., Lomedico, P. T., & Ju, G. (1984) · Nature · doi:10.1038/307241a0 · Cited by 368
Ghattas, I. R., Sanes, J. R., & Majors, J. E. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.12.5848-5859.1991 · Cited by 330
Huez, I., Créancier, L., Audigier, S., Gensac, M.-C., Prats, A., & Prats, H. (1998) · Molecular and Cellular Biology · doi:10.1128/mcb.18.11.6178 · Cited by 308
Rees, S., Coote, J., Stables, J., Goodson, S., Harris, S., & Lee, M. (1996) · BioTechniques · doi:10.2144/96201st05 · Cited by 299
Effects of intercistronic length on the efficiency of reinitiation by eucaryotic ribosomes.
Kozak, M. (1987) · Molecular and Cellular Biology · doi:10.1128/mcb.7.10.3438 · Cited by 278
Ghattas, I. R., Sanes, J. R., & Majors, J. E. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.12.5848 · Cited by 239
Borman, A. M., Le Mercier, P., Girard, M., & Kean, K. M. (1997) · Nucleic Acids Research · doi:10.1093/nar/25.5.925 · Cited by 238
Role for highly regulated rep gene expression in adeno-associated virus vector production
Li, J., Samulski, R. J., & Xiao, X. (1997) · Journal of Virology · doi:10.1128/jvi.71.7.5236-5243.1997 · Cited by 227
Morgan, R. A., Couture, L. A., Elroy‐Stein, O., Ragheb, J. A., Moss, B., & Anderson, W. F. (1992) · Nucleic Acids Research · doi:10.1093/nar/20.6.1293 · Cited by 204
Adam, M. A., Ramesh, N., Miller, A. D., & Osborne, W. R. A. (1991) · Journal of Virology · doi:10.1128/jvi.65.9.4985-4990.1991 · Cited by 196
Internal ribosome entry sites and dicistronic RNAs in mammalian transgenesis
Mountford, P. S., & Smith, A. G. (1995) · Trends in Genetics · doi:10.1016/s0168-9525(00)89040-x · Cited by 171
Quantitative analysis of gene suppression in integrated retrovirus vectors.
Emerman, M., & Temin, H. M. (1986) · Molecular and Cellular Biology · doi:10.1128/mcb.6.3.792 · Cited by 152
Borman, A. M., Bailly, J., Girard, M. P., & Kean, K. M. (1995) · Nucleic Acids Research · doi:10.1093/nar/23.18.3656 · Cited by 152
Chen, L., Chen, D., Block, E., O’Donnell, M. A., Kufe, D. W., & Clinton, S. K. (1997) · The Journal of Immunology · doi:10.4049/jimmunol.159.1.351 · Cited by 144
Okada, H., Giezeman‐Smits, K. M., Tahara, H., Attanucci, J., Fellows, W. K., Lotze, M. T., Chambers, W. H., & Bozik, M. E. (1999) · Gene Therapy · doi:10.1038/sj.gt.3300798 · Cited by 92
Gene therapy against an experimental glioma using adeno-associated virus vectors.
Okada, H., Miyamura, K., Itoh, T., Hagiwara, M., Wakabayashi, T., Mizuno, M., Colosi, P., Kurtzman, G. J., & Yoshida, J. (1996) · PubMed · Cited by 86
Efficient Gene Transfer into Mammalian Cells Using Fusogenic Liposome
Mizuguchi, H., Nakagawa, T., Nakanishi, M., Imazu, S., Nakagawa, S., & Mayumi, T. (1996) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1996.0070 · Cited by 83
Self-citationSharma, S. P., Miller, P. W., Stolina, M., Zhu, L., Huang, M., Paul, R. W., & Dubinett, S. M. (1997) · Gene Therapy · doi:10.1038/sj.gt.3300531 · Cited by 66
Pizzato, M., Franchin, E., Calvi, P., Boschetto, R., Colombo, M. P., Ferrini, S., & Palù, G. (1998) · Gene Therapy · doi:10.1038/sj.gt.3300670 · Cited by 47
Mizuguchi, H., Nakagawa, T., Toyosawa, S., Nakanishi, M., Imazu, S., Nakanishi, T., Tsutsumi, Y., Nakagawa, S., Hayakawa, T., Ijuhin, N., et al. (1998) · PubMed · Cited by 44
Self-citationA novel dicistronic AAV vector using a short IRES segment derived from hepatitis C virus genome
Urabe, M., Hasumi, Y., Ogasawara, Y., Matsushita, T., Kamoshita, N., Nomoto, A., Colosi, P., Kurtzman, G. J., Tobita, K., & Ozawa, K. (1997) · Gene · doi:10.1016/s0378-1119(97)00412-5 · Cited by 35
Fan, L., Drew, J., Dunckley, M. G., Owen, J., & Dickson, G. (1998) · Gene Therapy · doi:10.1038/sj.gt.3300746 · Cited by 34
Mizuguchi, H., Nakagawa, T., Morioka, Y., Imazu, S., Nakanishi, M., Kondo, T., Hayakawa, T., & Mayumi, T. (1997) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1997.6568 · Cited by 23
Self-citationHavenga, M. J. E., Vogels, R., Braakman, E., Kroos, N., Valerio, D., Hagenbeek, A., & van Es, H. H. G. (1998) · Gene · doi:10.1016/s0378-1119(98)00453-3 · Cited by 7
Flagged first, then by citations
Cited by
Sorted by citations
Engineering strategies to overcome the current roadblocks in CAR T cell therapy
Rafiq, S., Hackett, C. S., & Brentjens, R. J. (2019) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-019-0297-y · Cited by 1,525
Szymczak, A. L., Workman, C. J., Wang, Y., Vignali, K. M., Dilioglou, S., Vanin, E. F., & Vignali, D. A. A. (2004) · Nature Biotechnology · doi:10.1038/nbt957 · Cited by 1,059
Weinberger, L. S., Burnett, J., Toettcher, J. E., Arkin, A. P., & Schaffer, D. V. (2005) · Cell · doi:10.1016/j.cell.2005.06.006 · Cited by 633
Dawson, P. A., Hubbert, M. L., Haywood, J., Craddock, A. L., Zerangue, N., Christian, W. V., & Ballatori, N. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m412752200 · Cited by 381
Stable antibody expression at therapeutic levels using the 2A peptide
Fang, J., Qian, J., Yi, S., Harding, T. C., Tu, G. H., VanRoey, M. J., & Jooss, K. U. (2005) · Nature Biotechnology · doi:10.1038/nbt1087 · Cited by 370
Metadata
- 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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review