Uta Müller-Kuller, Mania Ackermann, Stephan Kolodziej, Christian Brendel, Jessica Fritsch, Nico Lachmann, Hana Kunkel, Jörn Lausen, Axel Schambach, Thomas Moritz, Manuel Grez
- Epigenetic silencing of transgene expression represents a major obstacle for the efficient genetic modification of multipotent and pluripotent stem cells. We and others have demonstrated that a 1.5 kb methylation-free CpG island from the human HNRPA2B1-CBX3 housekeeping genes (A2UCOE) effectively prevents transgene silencing and variegation in cell lines, multipotent and pluripotent stem cells, and their differentiated progeny. However, the bidirectional promoter activity of this element may disturb expression of neighboring genes. Furthermore, the epigenetic basis underlying the anti-silencing effect of the UCOE on juxtaposed promoters has been only partially explored. In this study we removed the HNRPA2B1 moiety from the A2UCOE and demonstrate efficient anti-silencing properties also for a minimal 0.7 kb element containing merely the CBX3 promoter. This DNA element largely prevents silencing of viral and tissue-specific promoters in multipotent and pluripotent stem cells. The protective activity of CBX3 was associated with reduced promoter CpG-methylation, decreased levels of repressive and increased levels of active histone marks. Moreover, the anti-silencing effect of CBX3 was locally restricted and when linked to tissue-specific promoters did not activate transcription in off target cells. Thus, CBX3 is a highly attractive element for sustained, tissue-specific and copy-number dependent transgene expression in vitro and in vivo.
MetadatenAuthor: | Uta Müller-KullerGND, Mania Ackermann, Stephan Kolodziej, Christian BrendelGND, Jessica Fritsch, Nico Lachmann, Hana Kunkel, Jörn Lausen, Axel Schambach, Thomas Moritz, Manuel GrezGND |
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URN: | urn:nbn:de:hebis:30:3-373853 |
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DOI: | https://doi.org/10.1093/nar/gkv019 |
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ISSN: | 1362-4962 |
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Pubmed Id: | https://pubmed.ncbi.nlm.nih.gov/25605798 |
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Parent Title (English): | Nucleic acids research : NAR |
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Publisher: | Oxford Univ. Press |
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Place of publication: | Oxford |
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Document Type: | Article |
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Language: | English |
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Date of Publication (online): | 2015/01/20 |
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Date of first Publication: | 2015/01/20 |
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Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
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Release Date: | 2015/06/19 |
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Volume: | 43 |
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Issue: | 3 |
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Page Number: | 16 |
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First Page: | 1577 |
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Last Page: | 1592 |
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Note: | Copyright © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact moc.puo@snoissimrep.slanruoj |
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HeBIS-PPN: | 369631358 |
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Institutes: | Medizin / Medizin |
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| Angeschlossene und kooperierende Institutionen / Georg-Speyer-Haus |
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Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
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| 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
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Sammlungen: | Universitätspublikationen |
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| Sammlung Biologie / Sondersammelgebiets-Volltexte |
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Licence (English): | Creative Commons - Namensnennung-Nicht kommerziell 4.0 |
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