MAR-mediated integration of plasmid vectors for in vivo gene transfer and regulation.

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Etat: Public
Version: de l'auteur
ID Serval
serval:BIB_B81D2BB6080C
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
MAR-mediated integration of plasmid vectors for in vivo gene transfer and regulation.
Périodique
BMC Molecular Biology
Auteur(s)
Puttini S., van Zwieten R.W., Saugy D., Lekka M., Hogger F., Ley D., Kulik A.J., Mermod N.
ISSN
1471-2199 (Electronic)
ISSN-L
1471-2199
Statut éditorial
Publié
Date de publication
2013
Volume
14
Pages
26
Langue
anglais
Résumé
BACKGROUND: The in vivo transfer of naked plasmid DNA into organs such as muscles is commonly used to assess the expression of prophylactic or therapeutic genes in animal disease models.
RESULTS: In this study, we devised vectors allowing a tight regulation of transgene expression in mice from such non-viral vectors using a doxycycline-controlled network of activator and repressor proteins. Using these vectors, we demonstrate proper physiological response as consequence of the induced expression of two therapeutically relevant proteins, namely erythropoietin and utrophin. Kinetic studies showed that the induction of transgene expression was only transient, unless epigenetic regulatory elements termed Matrix Attachment Regions, or MAR, were inserted upstream of the regulated promoters. Using episomal plasmid rescue and quantitative PCR assays, we observed that similar amounts of plasmids remained in muscles after electrotransfer with or without MAR elements, but that a significant portion had integrated into the muscle fiber chromosomes. Interestingly, the MAR elements were found to promote plasmid genomic integration but to oppose silencing effects in vivo, thereby mediating long-term expression.
CONCLUSIONS: This study thus elucidates some of the determinants of transient or sustained expression from the use of non-viral regulated vectors in vivo.
Pubmed
Web of science
Open Access
Oui
Création de la notice
28/01/2014 12:37
Dernière modification de la notice
20/08/2019 15:26
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