Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE)
Abstract Background DNA transposon-based vectors are effective nonviral tools for gene therapy and genetic engineering of cells. However, promoter DNA methylation and a near-random integration profile, which can result in transgene integration into heterochromatin, renders such vectors vulnerable to...
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| Format: | Article |
| Language: | English |
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BMC
2019-11-01
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| Series: | BMC Biotechnology |
| Online Access: | http://link.springer.com/article/10.1186/s12896-019-0570-2 |
| _version_ | 1818523348899987456 |
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| author | Kristian Alsbjerg Skipper Anne Kruse Hollensen Michael N. Antoniou Jacob Giehm Mikkelsen |
| author_facet | Kristian Alsbjerg Skipper Anne Kruse Hollensen Michael N. Antoniou Jacob Giehm Mikkelsen |
| author_sort | Kristian Alsbjerg Skipper |
| collection | DOAJ |
| description | Abstract Background DNA transposon-based vectors are effective nonviral tools for gene therapy and genetic engineering of cells. However, promoter DNA methylation and a near-random integration profile, which can result in transgene integration into heterochromatin, renders such vectors vulnerable to transcriptional repression. Therefore, to secure persistent transgene expression it may be necessary to protect transposon-embedded transgenes with anti-transcriptional silencing elements. Results We compare four different protective strategies in CHO-K1 cells. Our findings show robust protection from silencing of transgene cassettes mediated by the ubiquitous chromatin-opening element (UCOE) derived from the HNRPA2B1-CBX3 locus. Using a bioinformatic approach, we define a shorter HNRPA2B1-CBX3 UCOE core fragment and demonstrate that this can robustly maintain transgene expression after extended passaging of CHO-K1 cells carrying DNA transposon vectors equipped with this protective feature. Conclusions Our findings contribute to the understanding of the mechanism of HNRPA2B1-CBX3 UCOE-based transgene protection and support the use of a correctly oriented core fragment of this UCOE for DNA transposon vector-based production of recombinant proteins in CHO-K1 cells. |
| first_indexed | 2024-12-11T05:43:53Z |
| format | Article |
| id | doaj.art-144b1e9f55574905a37cb7eb483f5a62 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-11T05:43:53Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-144b1e9f55574905a37cb7eb483f5a622022-12-22T01:19:02ZengBMCBMC Biotechnology1472-67502019-11-0119111110.1186/s12896-019-0570-2Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE)Kristian Alsbjerg Skipper0Anne Kruse Hollensen1Michael N. Antoniou2Jacob Giehm Mikkelsen3Department of Biomedicine, HEALTH, Aarhus UniversityDepartment of Biomedicine, HEALTH, Aarhus UniversityGene Expression and Therapy Group, King’s College London, Faculty of Life Sciences & Medicine, Department of Medical and Molecular GeneticsDepartment of Biomedicine, HEALTH, Aarhus UniversityAbstract Background DNA transposon-based vectors are effective nonviral tools for gene therapy and genetic engineering of cells. However, promoter DNA methylation and a near-random integration profile, which can result in transgene integration into heterochromatin, renders such vectors vulnerable to transcriptional repression. Therefore, to secure persistent transgene expression it may be necessary to protect transposon-embedded transgenes with anti-transcriptional silencing elements. Results We compare four different protective strategies in CHO-K1 cells. Our findings show robust protection from silencing of transgene cassettes mediated by the ubiquitous chromatin-opening element (UCOE) derived from the HNRPA2B1-CBX3 locus. Using a bioinformatic approach, we define a shorter HNRPA2B1-CBX3 UCOE core fragment and demonstrate that this can robustly maintain transgene expression after extended passaging of CHO-K1 cells carrying DNA transposon vectors equipped with this protective feature. Conclusions Our findings contribute to the understanding of the mechanism of HNRPA2B1-CBX3 UCOE-based transgene protection and support the use of a correctly oriented core fragment of this UCOE for DNA transposon vector-based production of recombinant proteins in CHO-K1 cells.http://link.springer.com/article/10.1186/s12896-019-0570-2 |
| spellingShingle | Kristian Alsbjerg Skipper Anne Kruse Hollensen Michael N. Antoniou Jacob Giehm Mikkelsen Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE) BMC Biotechnology |
| title | Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE) |
| title_full | Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE) |
| title_fullStr | Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE) |
| title_full_unstemmed | Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE) |
| title_short | Sustained transgene expression from sleeping beauty DNA transposons containing a core fragment of the HNRPA2B1-CBX3 ubiquitous chromatin opening element (UCOE) |
| title_sort | sustained transgene expression from sleeping beauty dna transposons containing a core fragment of the hnrpa2b1 cbx3 ubiquitous chromatin opening element ucoe |
| url | http://link.springer.com/article/10.1186/s12896-019-0570-2 |
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