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Identification of anti-repressor elements that confer high and stable protein production in mammalian cells

Nature Biotechnology volume 21, pages 553–558 (2003)Cite this article

A Corrigendum to this article was published on 01 July 2003

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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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Figure 1: A genetic screen to identify DNA elements that block chromatin-associated repressors.
Figure 2: Anti-repressor elements are not enhancers, but they do block enhancers.
Figure 3: Anti-repressor elements confer more predictable and higher protein expression.
Figure 4: Anti-repressor elements confer stable protein expression in a copy number–dependent fashion.
Figure 5: Anti-repressor elements are highly conserved noncoding DNA.

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  • 14 April 2003

    This error has been corrected for the HTML (footnote, amended PDF until issue live) and the print versions of the article.

Notes

  1. *Note: In the version of this article originally published online, the last two sentences of the first paragraph of Results are incorrect. The correct text should read: "Four independent screens were performed using either LexA-HPC2 or LexA-HP1 as repressor. This resulted in the recovery of 65 pSelect plasmids that conveyed survival (Fig. 1B)."

    This error has been corrected for the HTML and the print versions of the article.

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Acknowledgements

We thank Valerie Dumay, Marike Feenstra, Thijs Hendrix, David Holmes, Mobien Kasiem, Ephie Kraneveld, Tim Segboer, and Johan van der Vlag for their involvement in the screens. P. Kwakman was sponsored by the Human Frontier Science Program (RG0039/1999-M).

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Authors and Affiliations

  1. Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, University of Amsterdam, Plantage Muidergracht 12, Amsterdam, 1018 TV

    Ted H.J. Kwaks, Phil Barnett, Wieger Hemrika, David P.E. Satijn, Paul Kwakman, Arle L. Kruckeberg & Arie P. Otte

  2. ChromaGenics BV, Plantage Muidergracht 12, Amsterdam, 1018 TV, The Netherlands

    Tjalling Siersma, Richard G.A.B. Sewalt, Janynke F. Brons, Rik van Blokland, Angèle Kelder & Arie P. Otte

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  1. Ted H.J. Kwaks
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Correspondence to Arie P. Otte.

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T.S., R.G.A.B.S, J.F.B., R.v.B., A.K., and A.P.O. are employed by ChromaGenics, a startup company that emerged from the Faculty of Sciences of the University of Amsterdam. ChromaGenics' mission is to create value through developing and commercializing proprietary technologies based on epigenetic gene regulation. ChromaGenics is within the compounds of the university and is financially supported by loans from the university and subsidies from the Ministry of Economical Affairs.

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Kwaks, T., Barnett, P., Hemrika, W. et al. Identification of anti-repressor elements that confer high and stable protein production in mammalian cells. Nat Biotechnol 21, 553–558 (2003). https://doi.org/10.1038/nbt814

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