Show simple item record

dc.contributor.authorKunkiel, Jessica
dc.contributor.authorGödecke, Natascha
dc.contributor.authorAckermann, Mania
dc.contributor.authorHoffmann, Dirk
dc.contributor.authorSchambach, Axel
dc.contributor.authorLachmann, Nico
dc.contributor.authorWirth, Dagmar
dc.contributor.authorMoritz, Thomas
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.
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
refterms.dateFOA2018-06-13T00:04:03Z
html.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.


Files in this item

Thumbnail
Name:
Kunkiel et al.pdf
Size:
4.340Mb
Format:
PDF
Description:
Open Access publication

This item appears in the following Collection(s)

Show simple item record

http://creativecommons.org/licenses/by-nc-sa/4.0/
Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-sa/4.0/