Genetic modification of dividing cells using episomally maintained S/MAR DNA vectors
The development of episomally maintained DNA vectors to genetically modify dividing cells efficiently and stably, without the risk of integration-mediated genotoxicity, should prove to be a valuable tool in genetic research. In this study, we demonstrate the utility of Scaffold/Matrix Attachment Reg...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2013-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253116301731 |
| _version_ | 1828155921423925248 |
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| author | Suet-Ping Wong Richard Paul Harbottle |
| author_facet | Suet-Ping Wong Richard Paul Harbottle |
| author_sort | Suet-Ping Wong |
| collection | DOAJ |
| description | The development of episomally maintained DNA vectors to genetically modify dividing cells efficiently and stably, without the risk of integration-mediated genotoxicity, should prove to be a valuable tool in genetic research. In this study, we demonstrate the utility of Scaffold/Matrix Attachment Region (S/MAR) DNA vectors to model the restoration of a functional wild-type copy of the gene folliculin (FLCN) implicated in the renal cancer Birt-Hogg-Dubé (BHD). Inactivation of FLCN has been shown to be involved in the development of sporadic renal neoplasia in BHD. S/MAR-modified BHD tumor cells (named UOK257-FS) show restored stable FLCN expression and have normalized downstream TGFβ signals. We demonstrate that UOK257-FS cells show a reduced growth rate in vitro and suppression of xenograft tumor development in vivo, compared with the original FLCN-null UOK257 cell line. In addition, we demonstrate that mTOR signaling in serum-starved FLCN-restored cells is differentially regulated compared with the FLCN-deficient cell. The novel UOK257-FS cell line will be useful for studying the signaling pathways affected in BHD pathogenesis. Significantly, this study demonstrates the suitability of S/MAR vectors to successfully model the functional expression of a therapeutic gene in a cancer cell line and will aid the identification of novel cancer markers for diagnosis and therapy. |
| first_indexed | 2024-04-11T23:01:36Z |
| format | Article |
| id | doaj.art-3b4ca08773e64d99a1cd4326fa0cd6d5 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-04-11T23:01:36Z |
| publishDate | 2013-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-3b4ca08773e64d99a1cd4326fa0cd6d52022-12-22T03:58:08ZengElsevierMolecular Therapy: Nucleic Acids2162-25312013-01-012C10.1038/mtna.2013.40Genetic modification of dividing cells using episomally maintained S/MAR DNA vectorsSuet-Ping Wong0Richard Paul Harbottle1Gene Therapy Group, Molecular Medicine, National Heart and Lung Institute, Imperial College London, London, UKGene Therapy Group, Molecular Medicine, National Heart and Lung Institute, Imperial College London, London, UKThe development of episomally maintained DNA vectors to genetically modify dividing cells efficiently and stably, without the risk of integration-mediated genotoxicity, should prove to be a valuable tool in genetic research. In this study, we demonstrate the utility of Scaffold/Matrix Attachment Region (S/MAR) DNA vectors to model the restoration of a functional wild-type copy of the gene folliculin (FLCN) implicated in the renal cancer Birt-Hogg-Dubé (BHD). Inactivation of FLCN has been shown to be involved in the development of sporadic renal neoplasia in BHD. S/MAR-modified BHD tumor cells (named UOK257-FS) show restored stable FLCN expression and have normalized downstream TGFβ signals. We demonstrate that UOK257-FS cells show a reduced growth rate in vitro and suppression of xenograft tumor development in vivo, compared with the original FLCN-null UOK257 cell line. In addition, we demonstrate that mTOR signaling in serum-starved FLCN-restored cells is differentially regulated compared with the FLCN-deficient cell. The novel UOK257-FS cell line will be useful for studying the signaling pathways affected in BHD pathogenesis. Significantly, this study demonstrates the suitability of S/MAR vectors to successfully model the functional expression of a therapeutic gene in a cancer cell line and will aid the identification of novel cancer markers for diagnosis and therapy.http://www.sciencedirect.com/science/article/pii/S2162253116301731 |
| spellingShingle | Suet-Ping Wong Richard Paul Harbottle Genetic modification of dividing cells using episomally maintained S/MAR DNA vectors Molecular Therapy: Nucleic Acids |
| title | Genetic modification of dividing cells using episomally maintained S/MAR DNA vectors |
| title_full | Genetic modification of dividing cells using episomally maintained S/MAR DNA vectors |
| title_fullStr | Genetic modification of dividing cells using episomally maintained S/MAR DNA vectors |
| title_full_unstemmed | Genetic modification of dividing cells using episomally maintained S/MAR DNA vectors |
| title_short | Genetic modification of dividing cells using episomally maintained S/MAR DNA vectors |
| title_sort | genetic modification of dividing cells using episomally maintained s mar dna vectors |
| url | http://www.sciencedirect.com/science/article/pii/S2162253116301731 |
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