The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function.

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Hdl Handle:
http://hdl.handle.net/10033/621057
Title:
The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function.
Authors:
Kunkiel, Jessica; Gödecke, Natascha; Ackermann, Mania; Hoffmann, Dirk; Schambach, Axel; Lachmann, Nico; Wirth, Dagmar; Moritz, Thomas
Abstract:
Suppression of therapeutic transgene expression from retroviral gene therapy vectors by epigenetic defence mechanisms represents a problem that is particularly encountered in pluripotent stem cells (PSCs) and their differentiated progeny. Transgene expression in these cells, however, can be stabilised by CpG-rich ubiquitous chromatin opening elements (UCOEs). In this context we recently demonstrated profound anti-silencing properties for the small (679 bp) CBX3-UCO element and we now confirmed this observation in the context of the defined murine chromosomal loci ROSA26 and TIGRE. Moreover, since the structural basis for the anti-silencing activity of UCOEs has remained poorly defined, we interrogated various CBX3 subfragments in the context of lentiviral vectors and murine PSCs. We demonstrated marked though distinct anti-silencing activity in the pluripotent state and during PSC-differentiation for several of the CBX3 subfragments. This activity was significantly correlated with CpG content as well as endogenous transcriptional activity. Interestingly, also a scrambled CBX3 version with preserved CpG-sites retained the anti-silencing activity despite the lack of endogenous promoter activity. Our data therefore highlight the importance of CpG-sites and transcriptional activity for UCOE functionality and suggest contributions from different mechanisms to the overall anti-silencing function of the CBX3 element.
Affiliation:
Helmholtz Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function. 2017, 7 (1):7919 Sci Rep
Journal:
Scientific reports
Issue Date:
11-Aug-2017
URI:
http://hdl.handle.net/10033/621057
DOI:
10.1038/s41598-017-04212-8
PubMed ID:
28801671
Type:
Article
Language:
en
ISSN:
2045-2322
Appears in Collections:
publications of the research group modell systems for infections and immunity (MSYS)

Full metadata record

DC FieldValue Language
dc.contributor.authorKunkiel, Jessicaen
dc.contributor.authorGödecke, Nataschaen
dc.contributor.authorAckermann, Maniaen
dc.contributor.authorHoffmann, Dirken
dc.contributor.authorSchambach, Axelen
dc.contributor.authorLachmann, Nicoen
dc.contributor.authorWirth, Dagmaren
dc.contributor.authorMoritz, Thomasen
dc.date.accessioned2017-08-16T11:44:18Z-
dc.date.available2017-08-16T11:44:18Z-
dc.date.issued2017-08-11-
dc.identifier.citationThe CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function. 2017, 7 (1):7919 Sci Repen
dc.identifier.issn2045-2322-
dc.identifier.pmid28801671-
dc.identifier.doi10.1038/s41598-017-04212-8-
dc.identifier.urihttp://hdl.handle.net/10033/621057-
dc.description.abstractSuppression of therapeutic transgene expression from retroviral gene therapy vectors by epigenetic defence mechanisms represents a problem that is particularly encountered in pluripotent stem cells (PSCs) and their differentiated progeny. Transgene expression in these cells, however, can be stabilised by CpG-rich ubiquitous chromatin opening elements (UCOEs). In this context we recently demonstrated profound anti-silencing properties for the small (679 bp) CBX3-UCO element and we now confirmed this observation in the context of the defined murine chromosomal loci ROSA26 and TIGRE. Moreover, since the structural basis for the anti-silencing activity of UCOEs has remained poorly defined, we interrogated various CBX3 subfragments in the context of lentiviral vectors and murine PSCs. We demonstrated marked though distinct anti-silencing activity in the pluripotent state and during PSC-differentiation for several of the CBX3 subfragments. This activity was significantly correlated with CpG content as well as endogenous transcriptional activity. Interestingly, also a scrambled CBX3 version with preserved CpG-sites retained the anti-silencing activity despite the lack of endogenous promoter activity. Our data therefore highlight the importance of CpG-sites and transcriptional activity for UCOE functionality and suggest contributions from different mechanisms to the overall anti-silencing function of the CBX3 element.en
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleThe CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalScientific reportsen

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