Identification of anti-repressor elements that confer high and stable protein production in mammalian cells
Abstract
The expression of transgenic proteins is often low and unstable over time, a problem that may be due to integration of the transgene in repressed chromatin. We developed a screening technology to identify genetic elements that efficiently counteract chromatin-associated repression. When these elements were used to flank a transgene, we observed a substantial increase in the number of mammalian cell colonies that expressed the transgenic protein. Expression of the shielded transgene was, in a copy number-dependent fashion, substantially higher than the expression of unprotected transgenes. Also, protein production remained stable over an extended time period. The DNA elements are small, not exceeding 2,100 base pairs (bp), and they are highly conserved between human and mouse, at both the functional and sequence levels. Our results demonstrate the existence of a class of genetic elements that can readily be applied to more efficient transgenic protein production in mammalian cells.
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STAR elements were discovered by a genomic screen of elements that would increase transgene expression.
“They were discovered by a genomic screen of elements that would increase transgene expression.(Kwaks et al., 2003;Sautter and Enenkel, 2005;Hoeksema et al., 2011;Van Blokland et al., 2011;Harraghy et al., 2015).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Kwaks et al., 2003
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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
- Kwaks, T. H. J., Barnett, P., Hemrika, W., Siersma, T., Sewalt, R. G. A. B., Satijn, D. P. E., Brons, J. F., van Blokland, R., Kwakman, P., Kruckeberg, A. L., Kelder, A., & Otte, A. P.
- Affiliations
- University of Amsterdam (NL)
- Corresponding author
- Otte, Arie P.
- Venue
- Nature Biotechnology
- Published
- 2003-04-07
- Volume
- 21
- Issue
- 5
- Pages
- 553-558
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W2017907556
- Cited by
- 126
- References
- 0
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 3 · 2023: 4 · 2022: 3 · 2021: 3 · 2020: 5 · 2019: 5 · 2018: 3 · 2017: 7 · 2016: 6 · 2015: 7 · 2014: 9 · 2013: 8 · 2012: 5
- FWCI
- 1.96
- Type
- journal article
- Language
- en
- Update notices
- erratum (2003-07-01)
- Open access
- closed
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- UvA-DARE (University of Amsterdam) (publishedVersion)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- Pure Amsterdam UMC (submittedVersion)
- 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 › Animal Genetics and Reproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- chromatin repression, transgene expression, transgene integration, mammalian cells, expression stability, screening technologies, human-mouse sequence conservation
- MeSH terms
- Animals, Chromatin, Cricetulus, Gene Expression Regulation, Cricetinae, Humans, Mammals, Molecular Sequence Data, Promoter Regions, Genetic, Proteins, Recombinant Proteins, Repressor Proteins, Protein Biosynthesis, Protein Engineering, CHO Cells, Transgenes, Gene Expression Profiling
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