共 41 条
Stringent rosiglitazone-dependent gene switch in muscle cells without effect on myogenic differentiation
被引:16
作者:

Tascou, S
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Gencell SAS, F-94408 Vitry Sur Seine, France Gencell SAS, F-94408 Vitry Sur Seine, France

Sorensen, TK
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Gencell SAS, F-94408 Vitry Sur Seine, France Gencell SAS, F-94408 Vitry Sur Seine, France

Glénat, V
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Gencell SAS, F-94408 Vitry Sur Seine, France Gencell SAS, F-94408 Vitry Sur Seine, France

Wang, MP
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机构:
Gencell SAS, F-94408 Vitry Sur Seine, France Gencell SAS, F-94408 Vitry Sur Seine, France

Lakich, MM
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Gencell SAS, F-94408 Vitry Sur Seine, France Gencell SAS, F-94408 Vitry Sur Seine, France

Darteil, R
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h-index: 0
机构:
Gencell SAS, F-94408 Vitry Sur Seine, France Gencell SAS, F-94408 Vitry Sur Seine, France

Vigne, E
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h-index: 0
机构:
Gencell SAS, F-94408 Vitry Sur Seine, France Gencell SAS, F-94408 Vitry Sur Seine, France

Thuillier, V
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机构:
Gencell SAS, F-94408 Vitry Sur Seine, France Gencell SAS, F-94408 Vitry Sur Seine, France
机构:
[1] Gencell SAS, F-94408 Vitry Sur Seine, France
关键词:
gene therapy;
PPAR gamma;
adipogenesis;
muscle;
plasmid;
scaffold/matrix attachment region;
D O I:
10.1016/j.ymthe.2004.02.013
中图分类号:
Q81 [生物工程学(生物技术)];
Q93 [微生物学];
学科分类号:
071005 ;
0836 ;
090102 ;
100705 ;
摘要:
We have developed a gene switch based on the human transcription factor peroxisome proliferator-activated receptor gamma (PPARgamma) and its activation by rosiglitazone. However, ectopic expression of PPARgamma has been demonstrated to convert myogenic cells into adipocyte-like cells and, more generally, may interfere with the physiology of the target tissue. Consequently we modified the DNA-binding specificity of PPARgamma, resulting in a transcription factor that we named PPAR*. We demonstrated by histological and molecular assessment of cell phenotype that the overexpression of PPAR* did not alter the myogenic differentiation program of G8 myoblasts. We showed that PPAR* does not transactivate promoters containing PPARgamma-responsive elements but transactivates promoters containing PPAR*-responsive elements that are at least 80% identical to a 20-bp consensus. We improved the rosiglitazone-dependent gene switch by tuning PPAR* expression with a scaffold/matrix attachment region and by expressing both PPAR* and the reporter gene under the control of PPAR*-responsive elements. Treatment of cultured murine muscle cells (myotubes) with rosiglitazone induced reporter gene expression from assay background up to the level attained by a CMV I/E promoter-enhancer. These results indicate the potential of the PPAR* gene switch for use in gene therapy applications.
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页码:637 / 649
页数:13
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