Improvement of in vivo transfer of plasmid DNA in muscle: comparison of electroporation versus ultrasound.
Plasmid-based gene delivery to muscle is a treatment strategy for many diseases with potential advantages above viral-based gene delivery methods, however, with a relative low transfection efficiency. We compared two physical methods - electroporation and ultrasound - that facilitate DNA uptake into cells. Mice (C57Bl/6) were injected intramuscular using plasmid DNA encoding an intracellular protein (p53) followed by electroporation or ultrasound. Then 48 hr after the injections the mice were sacrificed. The parameter for transfection efficiency was the area of muscle expressing the transgene. The p53 expression plasmid showed a 36-fold increase (p = 0.015) in transfection efficiency with electroporation compared to ultrasound. Compared with ultrasound, electroporation significantly improves transfection efficiency of naked plasmid DNA transfer into skeletal muscle.
Kusumanto
Yoka H
YH
Department of Medical Oncology, University of Groningen and University Medical Center Groningen, Groningen, The Netherlands.
Mulder
Nanno H
NH
Dam
Wendy A
WA
Losen
Mario
M
Losen
Mario H
MH
De Baets
Marc H
MH
Meijer
Coby
C
Hospers
Geke A P
GA
D000818:Animals; D004247:DNA / Q000235:genetics*; D018274:Electroporation / Q000379:methods; D005260:Female; D007150:Immunohistochemistry; D007273:Injections, Intramuscular; D051379:Mice; D008810:Mice, Inbred C57BL; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000378:metabolism; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D014162:Transfection / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism; D014465:Ultrasonics
D011993:Recombinant Fusion Proteins; D016159:Tumor Suppressor Protein p53; D004247:DNA