Minimal size MIDGE vectors improve transgene expression in vivo.
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.
Schakowski
Frank
F
Department of Internal Medicine I, University of Bonn, Germany.
Gorschlüter
Marcus
M
Buttgereit
Peter
P
Märten
Angela
A
Lilienfeld-Toal
Marie V
MV
Junghans
Claas
C
Schroff
Matthias
M
König-Merediz
Sven A
SA
Ziske
Carsten
C
Strehl
John
J
Sauerbruch
Tilman
T
Wittig
Burghardt
B
Schmidt-Wolf
Ingo G H
IG
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
D008156:Luciferases