title: Minimal size MIDGE vectors improve transgene expression in vivo.
abstract: Viral and plasmid vectors may cause immunological side-effects resulting from the expression of therapeutically unwanted genes and from CpG motifs contained in their sequence. A new vector type for minimalistic, immunological-defined gene expression (MIDGE) may overcome these problems. MIDGE is a minimal size gene transfer unit consisting of the expression cassette, including promotor, gene and RNA-stabilizing sequences, flanked by two short hairpin oligonucleotide sequences. DNA not encoding the desired gene is reduced to a minimum. To compare transfection efficiencies in vivo hydrodynamics-based, systemic transfection was performed in BALB/c mice with MIDGE vectors and corresponding plasmids. The transfection efficiencies of the MIDGE vectors as measured by luciferase expression were significantly higher in liver (2.5-fold), lung (3.5-fold), kidneys (3.9-fold) and heart (17-fold) as compared to plasmids. The mean numbers of MIDGE vector molecules per cell as measured by quantitative PCR were also significantly higher. These advantages suggest the preferential use of this new vector type for clinical gene therapy studies.
authors.affiliation: Department of Internal Medicine I, University of Bonn, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Schakowski
authors.forename: Marcus
authors.initials: M
authors.lastname: Gorschlüter
authors.forename: Peter
authors.initials: P
authors.lastname: Buttgereit
authors.forename: Angela
authors.initials: A
authors.lastname: Märten
authors.forename: Marie V
authors.initials: MV
authors.lastname: Lilienfeld-Toal
authors.forename: Claas
authors.initials: C
authors.lastname: Junghans
authors.forename: Matthias
authors.initials: M
authors.lastname: Schroff
authors.forename: Sven A
authors.initials: SA
authors.lastname: König-Merediz
authors.forename: Carsten
authors.initials: C
authors.lastname: Ziske
authors.forename: John
authors.initials: J
authors.lastname: Strehl
authors.forename: Tilman
authors.initials: T
authors.lastname: Sauerbruch
authors.forename: Burghardt
authors.initials: B
authors.lastname: Wittig
authors.forename: Ingo G H
authors.initials: IG
authors.lastname: Schmidt-Wolf
mesh_terms: D000818:Animals; D018899:CpG Islands; D005822:Genetic Vectors; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D009928:Organ Specificity; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014162:Transfection / Q000379:methods*; D019076:Transgenes
chemical_list: D008156:Luciferases