The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function
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...
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2017
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oai:doaj.org-article:39b1333cdd3f49e5b6f13a3b49b6f0d62021-12-02T12:31:50ZThe CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function10.1038/s41598-017-04212-82045-2322https://doaj.org/article/39b1333cdd3f49e5b6f13a3b49b6f0d62017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04212-8https://doaj.org/toc/2045-2322Abstract 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.Jessica KunkielNatascha GödeckeMania AckermannDirk HoffmannAxel SchambachNico LachmannDagmar WirthThomas MoritzNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017) |
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Medicine R Science Q Jessica Kunkiel Natascha Gödecke Mania Ackermann Dirk Hoffmann Axel Schambach Nico Lachmann Dagmar Wirth Thomas Moritz The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function |
| description |
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. |
| format |
article |
| author |
Jessica Kunkiel Natascha Gödecke Mania Ackermann Dirk Hoffmann Axel Schambach Nico Lachmann Dagmar Wirth Thomas Moritz |
| author_facet |
Jessica Kunkiel Natascha Gödecke Mania Ackermann Dirk Hoffmann Axel Schambach Nico Lachmann Dagmar Wirth Thomas Moritz |
| author_sort |
Jessica Kunkiel |
| title |
The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function |
| title_short |
The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function |
| title_full |
The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function |
| title_fullStr |
The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function |
| title_full_unstemmed |
The CpG-sites of the CBX3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function |
| title_sort |
cpg-sites of the cbx3 ubiquitous chromatin opening element are critical structural determinants for the anti-silencing function |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/39b1333cdd3f49e5b6f13a3b49b6f0d6 |
| work_keys_str_mv |
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1718394270459625472 |