pEPito: a significantly improved non-viral episomal expression vector for mammalian cells
<p>Abstract</p> <p>Background</p> <p>The episomal replication of the prototype vector pEPI-1 depends on a transcription unit starting from the constitutively expressed <it>Cytomegalovirus </it>immediate early promoter (CMV-IEP) and directed into a 2000 bp lo...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2010-03-01
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| Series: | BMC Biotechnology |
| Online Access: | http://www.biomedcentral.com/1472-6750/10/20 |
| _version_ | 1828259343233974272 |
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| author | Ogris Manfred Lipps Hans J Harbottle Richard P Wong Suet-Ping Argyros Orestis Haase Rudolf Magnusson Terese Pinto Maria Haas Jürgen Baiker Armin |
| author_facet | Ogris Manfred Lipps Hans J Harbottle Richard P Wong Suet-Ping Argyros Orestis Haase Rudolf Magnusson Terese Pinto Maria Haas Jürgen Baiker Armin |
| author_sort | Ogris Manfred |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The episomal replication of the prototype vector pEPI-1 depends on a transcription unit starting from the constitutively expressed <it>Cytomegalovirus </it>immediate early promoter (CMV-IEP) and directed into a 2000 bp long <it>matrix attachment region sequence </it>(MARS) derived from the human β-interferon gene. The original pEPI-1 vector contains two mammalian transcription units and a total of 305 CpG islands, which are located predominantly within the vector elements necessary for bacterial propagation and known to be counterproductive for persistent long-term transgene expression.</p> <p>Results</p> <p>Here, we report the development of a novel vector pEPito, which is derived from the pEPI-1 plasmid replicon but has considerably improved efficacy both <it>in vitro </it>and <it>in vivo</it>. The pEPito vector is significantly reduced in size, contains only one transcription unit and 60% less CpG motives in comparison to pEPI-1. It exhibits major advantages compared to the original pEPI-1 plasmid, including higher transgene expression levels and increased colony-forming efficiencies <it>in vitro</it>, as well as more persistent transgene expression profiles <it>in vivo</it>. The performance of pEPito-based vectors was further improved by replacing the CMV-IEP with the <it>human CMV enhancer/human elongation factor 1 alpha promoter </it>(hCMV/EF1P) element that is known to be less affected by epigenetic silencing events.</p> <p>Conclusions</p> <p>The novel vector pEPito can be considered suitable as an improved vector for biotechnological applications <it>in vitro </it>and for non-viral gene delivery <it>in vivo</it>.</p> |
| first_indexed | 2024-04-13T03:07:19Z |
| format | Article |
| id | doaj.art-43c95a8b7eb741b6bad7097d49fa8c27 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-04-13T03:07:19Z |
| publishDate | 2010-03-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-43c95a8b7eb741b6bad7097d49fa8c272022-12-22T03:05:12ZengBMCBMC Biotechnology1472-67502010-03-011012010.1186/1472-6750-10-20pEPito: a significantly improved non-viral episomal expression vector for mammalian cellsOgris ManfredLipps Hans JHarbottle Richard PWong Suet-PingArgyros OrestisHaase RudolfMagnusson TeresePinto MariaHaas JürgenBaiker Armin<p>Abstract</p> <p>Background</p> <p>The episomal replication of the prototype vector pEPI-1 depends on a transcription unit starting from the constitutively expressed <it>Cytomegalovirus </it>immediate early promoter (CMV-IEP) and directed into a 2000 bp long <it>matrix attachment region sequence </it>(MARS) derived from the human β-interferon gene. The original pEPI-1 vector contains two mammalian transcription units and a total of 305 CpG islands, which are located predominantly within the vector elements necessary for bacterial propagation and known to be counterproductive for persistent long-term transgene expression.</p> <p>Results</p> <p>Here, we report the development of a novel vector pEPito, which is derived from the pEPI-1 plasmid replicon but has considerably improved efficacy both <it>in vitro </it>and <it>in vivo</it>. The pEPito vector is significantly reduced in size, contains only one transcription unit and 60% less CpG motives in comparison to pEPI-1. It exhibits major advantages compared to the original pEPI-1 plasmid, including higher transgene expression levels and increased colony-forming efficiencies <it>in vitro</it>, as well as more persistent transgene expression profiles <it>in vivo</it>. The performance of pEPito-based vectors was further improved by replacing the CMV-IEP with the <it>human CMV enhancer/human elongation factor 1 alpha promoter </it>(hCMV/EF1P) element that is known to be less affected by epigenetic silencing events.</p> <p>Conclusions</p> <p>The novel vector pEPito can be considered suitable as an improved vector for biotechnological applications <it>in vitro </it>and for non-viral gene delivery <it>in vivo</it>.</p>http://www.biomedcentral.com/1472-6750/10/20 |
| spellingShingle | Ogris Manfred Lipps Hans J Harbottle Richard P Wong Suet-Ping Argyros Orestis Haase Rudolf Magnusson Terese Pinto Maria Haas Jürgen Baiker Armin pEPito: a significantly improved non-viral episomal expression vector for mammalian cells BMC Biotechnology |
| title | pEPito: a significantly improved non-viral episomal expression vector for mammalian cells |
| title_full | pEPito: a significantly improved non-viral episomal expression vector for mammalian cells |
| title_fullStr | pEPito: a significantly improved non-viral episomal expression vector for mammalian cells |
| title_full_unstemmed | pEPito: a significantly improved non-viral episomal expression vector for mammalian cells |
| title_short | pEPito: a significantly improved non-viral episomal expression vector for mammalian cells |
| title_sort | pepito a significantly improved non viral episomal expression vector for mammalian cells |
| url | http://www.biomedcentral.com/1472-6750/10/20 |
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