CpG-island fragments from the <it>HNRPA2B1</it>/<it>CBX3 </it>genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells
<p>Abstract</p> <p>Background</p> <p>The <it>hCMV </it>promoter is very commonly used for high level expression of transgenes in mammalian cells, but its utility is hindered by transcriptional silencing. Large genomic fragments incorporating the CpG island r...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2005-06-01
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| Series: | BMC Biotechnology |
| Online Access: | http://www.biomedcentral.com/1472-6750/5/17 |
| _version_ | 1818917506698444800 |
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| author | Irvine Alistair Antoniou Michael Simpson David Griffiths Mark Mulcahy Tony Mustoe Tracey Williams Steven Mountain Andrew Crombie Robert |
| author_facet | Irvine Alistair Antoniou Michael Simpson David Griffiths Mark Mulcahy Tony Mustoe Tracey Williams Steven Mountain Andrew Crombie Robert |
| author_sort | Irvine Alistair |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The <it>hCMV </it>promoter is very commonly used for high level expression of transgenes in mammalian cells, but its utility is hindered by transcriptional silencing. Large genomic fragments incorporating the CpG island region of the <it>HNRPA2B1 </it>locus are resistant to transcriptional silencing.</p> <p>Results </p> <p>In this report we describe studies on the use of a novel series of vectors combining the <it>HNRPA2B1 </it>CpG island with the hCMV promoter for expression of transgenes in CHO-K1 cells. We show that the CpG island gives at least twenty-fold increases in the levels of EGFP and EPO observed in pools of transfectants, and that transgene expression levels remain high in such pools for more than 100 generations. These novel vectors also allow facile isolation of clonal CHO-K1 cell lines showing stable, high-level transgene expression.</p> <p>Conclusion</p> <p>Vectors incorporating the <it>hnRPA2B1 </it>CpG island give major benefits in transgene expression from the <it>hCMV </it>promoter, including substantial improvements in the level and stability of expression. The utility of these vectors for the improved production of recombinant proteins in CHO cells has been demonstrated.</p> |
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| format | Article |
| id | doaj.art-59e435f88c1e4c07930d2aebce518606 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-20T00:35:09Z |
| publishDate | 2005-06-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-59e435f88c1e4c07930d2aebce5186062022-12-21T19:59:47ZengBMCBMC Biotechnology1472-67502005-06-01511710.1186/1472-6750-5-17CpG-island fragments from the <it>HNRPA2B1</it>/<it>CBX3 </it>genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cellsIrvine AlistairAntoniou MichaelSimpson DavidGriffiths MarkMulcahy TonyMustoe TraceyWilliams StevenMountain AndrewCrombie Robert<p>Abstract</p> <p>Background</p> <p>The <it>hCMV </it>promoter is very commonly used for high level expression of transgenes in mammalian cells, but its utility is hindered by transcriptional silencing. Large genomic fragments incorporating the CpG island region of the <it>HNRPA2B1 </it>locus are resistant to transcriptional silencing.</p> <p>Results </p> <p>In this report we describe studies on the use of a novel series of vectors combining the <it>HNRPA2B1 </it>CpG island with the hCMV promoter for expression of transgenes in CHO-K1 cells. We show that the CpG island gives at least twenty-fold increases in the levels of EGFP and EPO observed in pools of transfectants, and that transgene expression levels remain high in such pools for more than 100 generations. These novel vectors also allow facile isolation of clonal CHO-K1 cell lines showing stable, high-level transgene expression.</p> <p>Conclusion</p> <p>Vectors incorporating the <it>hnRPA2B1 </it>CpG island give major benefits in transgene expression from the <it>hCMV </it>promoter, including substantial improvements in the level and stability of expression. The utility of these vectors for the improved production of recombinant proteins in CHO cells has been demonstrated.</p>http://www.biomedcentral.com/1472-6750/5/17 |
| spellingShingle | Irvine Alistair Antoniou Michael Simpson David Griffiths Mark Mulcahy Tony Mustoe Tracey Williams Steven Mountain Andrew Crombie Robert CpG-island fragments from the <it>HNRPA2B1</it>/<it>CBX3 </it>genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells BMC Biotechnology |
| title | CpG-island fragments from the <it>HNRPA2B1</it>/<it>CBX3 </it>genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells |
| title_full | CpG-island fragments from the <it>HNRPA2B1</it>/<it>CBX3 </it>genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells |
| title_fullStr | CpG-island fragments from the <it>HNRPA2B1</it>/<it>CBX3 </it>genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells |
| title_full_unstemmed | CpG-island fragments from the <it>HNRPA2B1</it>/<it>CBX3 </it>genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells |
| title_short | CpG-island fragments from the <it>HNRPA2B1</it>/<it>CBX3 </it>genomic locus reduce silencing and enhance transgene expression from the hCMV promoter/enhancer in mammalian cells |
| title_sort | cpg island fragments from the it hnrpa2b1 it it cbx3 it genomic locus reduce silencing and enhance transgene expression from the hcmv promoter enhancer in mammalian cells |
| url | http://www.biomedcentral.com/1472-6750/5/17 |
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