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

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Version: author
Serval ID
serval:BIB_B81D2BB6080C
Type
Article: article from journal or magazin.
Collection
Publications
Title
MAR-mediated integration of plasmid vectors for in vivo gene transfer and regulation.
Journal
BMC Molecular Biology
Author(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
Publication state
Published
Issued date
2013
Volume
14
Pages
26
Language
english
Abstract
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
Yes
Create date
28/01/2014 13:37
Last modification date
20/08/2019 16:26
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