CpG-island fragments from the HNRPA2B1/CBX3genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells
Open accessAbstract
Background: The hCMV promoter is very commonly used for high level expression of transgenes in mammalian cells, but its utility is hindered by transcriptional silencing. Large genomic fragments incorporating the CpG island region of the HNRPA2B1 locus are resistant to transcriptional silencing. Results: In this report we describe studies on the use of a novel series of vectors combining the HNRPA2B1 CpG island with the hCMV promoter for expression of transgenes in CHO-K1 cells. We show that the CpG island gives at least twenty-fold increases in the levels of EGFP and EPO observed in pools of transfectants, and that transgene expression levels remain high in such pools for more than 100 generations. These novel vectors also allow facile isolation of clonal CHO-K1 cell lines showing stable, high-level transgene expression. Conclusion: Vectors incorporating the hnRPA2B1 CpG island give major benefits in transgene expression from the hCMV promoter, including substantial improvements in the level and stability of expression. The utility of these vectors for the improved production of recombinant proteins in CHO cells has been demonstrated.
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MAR and UCOE helped decrease the variability of expression between different clones and contributed to maintaining an active transcriptional state for better transgene expression.
“It was also demonstrated that MAR and UCOE helped decrease the variability of expression between the different clones and contributed in maintaining an active transcriptional state for better transgene expression (Williams et al., 2005;Galbete et al., 2009;Harraghy et al., 2011).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Williams et al., 2005
The Results and Discussion report that the CpG-island vectors increased the proportion of expressing clones, and the Conclusion says the findings are consistent with UCOEs maintaining a more open chromatin domain. But the paper does not demonstrate reduced expression variability between clones with the vectors studied; the Conclusion says they showed greater variability than the larger CpG-island construct used in earlier work. This paper studies UCOEs, not MARs. The indexed copy provides section labels but no page numbers. (LLM, conf=0.86)
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Research works printed but not linked: 9
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- Antoniou M Harland L Mustoe T Williams S Holdstock J Yague E Mulcahy T Griffiths M Edwards S Ioannou PA Mountain A Crombie R Transgenes encompassing dual-promoter CpG islands from the human TBP and HNRPA2B1 loci are resistant to heterochromatin-mediated silencing Genomics 2003 82 269 279 12906852 10.1016/S0888-7543(03)00107-1 · DOI
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Therapeutic glycoprotein production in mammalian cells
Lalonde, M.-E., & Durocher, Y. (2017) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2017.04.028 · Cited by 296 · see citing works
Harraghy, N., Calabrese, D., Fisch, I., Girod, P., LeFourn, V., Regamey, A., & Mermod, N. (2015) · Biotechnology Journal · doi:10.1002/biot.201400649 · Cited by 32 · see citing works
- Authors
- Williams, S., Mustoe, T., Mulcahy, T., Griffiths, M., Simpson, D., Antoniou, M., Irvine, A., Mountain, A., & Crombie, R.
- Affiliations
- Keele University (GB)
- King's College London (GB)
- Corresponding author
- Mountain, Andrew
- Venue
- BMC Biotechnology
- Published
- 2005-06-03
- Volume
- 5
- Issue
- 1
- Pages
- 17-17
- ISSN
- 1472-6750 (ISSN-L)
- PMC
- PMC1175082
- OpenAlex
- W1760112062
- Cited by
- 151
- References
- 0
- Citations by year
- 2026: 3 · 2025: 7 · 2024: 2 · 2023: 5 · 2022: 3 · 2021: 3 · 2020: 3 · 2019: 8 · 2018: 5 · 2017: 10 · 2016: 6 · 2015: 13 · 2014: 11 · 2013: 12 · 2012: 11
- FWCI
- 1.51
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- BMC Biotechnology (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Directorate for Biological Sciences
- Medical Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Keywords
- CpG islands, CBX3, transcriptional silencing, transgene expression, CHO-K1 cells, recombinant protein expression, expression vectors, enhanced green fluorescent protein, erythropoietin
- MeSH terms
- Animals, Biotechnology, Chromosomal Proteins, Non-Histone, Chromosome Mapping, Cytomegalovirus, Enhancer Elements, Genetic, Flow Cytometry, Gene Expression Regulation, Genetic Techniques, Genetic Vectors, Cricetinae, Models, Genetic, Promoter Regions, Genetic, Recombinant Proteins, Transcription, Genetic, Transfection, Gene Expression, CHO Cells, Genes, Reporter, CpG Islands, Transgenes, Gene Silencing, Green Fluorescent Proteins
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