Fiche du document
2013
- doi: 10.1186/1471-2199-14-26
- issn: 1471-2199
Ce document est lié à :
info:eu-repo/semantics/altIdentifier/doi/10.1186/1471-2199-14-26
Ce document est lié à :
info:eu-repo/semantics/altIdentifier/pmid/24295286
Ce document est lié à :
info:eu-repo/semantics/altIdentifier/eissn/1471-2199
Ce document est lié à :
info:eu-repo/semantics/altIdentifier/urn/urn:nbn:ch:serval-BIB_B81D2BB6080C7
info:eu-repo/semantics/openAccess , Copying allowed only for non-profit organizations , https://serval.unil.ch/disclaimer
Sujets proches
Expressive behaviorCiter ce document
S. Puttini et al., « MAR-mediated integration of plasmid vectors for in vivo gene transfer and regulation. », Serveur académique Lausannois, ID : 10.1186/1471-2199-14-26
Métriques
Partage / Export
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.