title: MAR-mediated integration of plasmid vectors for in vivo gene transfer and regulation.
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.
authors.affiliation: Institute of Biotechnology, University of Lausanne, Lausanne, Switzerland. nicolas.mermod@unil.ch.
authors.forename: Stefania
authors.initials: S
authors.lastname: Puttini
authors.forename: Ruthger W
authors.initials: RW
authors.lastname: van Zwieten
authors.forename: Damien
authors.initials: D
authors.lastname: Saugy
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Lekka
authors.forename: Florence
authors.initials: F
authors.lastname: Hogger
authors.forename: Deborah
authors.initials: D
authors.lastname: Ley
authors.forename: Andrzej J
authors.initials: AJ
authors.lastname: Kulik
authors.forename: Nicolas
authors.initials: N
authors.lastname: Mermod
mesh_terms: D000818:Animals; D004318:Doxycycline / Q000494:pharmacology; D044127:Epigenesis, Genetic; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D007700:Kinetics; D045170:Matrix Attachment Regions; D051379:Mice; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D009132:Muscles / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D019076:Transgenes; D049411:Utrophin / Q000235:genetics / Q000378:metabolism
chemical_list: D049411:Utrophin; D004921:Erythropoietin; D004318:Doxycycline