In vitro gene transfer by electrosonoporation.
Among the nonviral methods for gene delivery in vitro, electroporation is simple, inexpensive and safe. To upregulate the expression level of transfected gene, we investigated the applicability of electrosonoporation. This approach consists of a combination of electric pulses and ultrasound assisted with gas microbubbles. Cells were first electroporated with plasmid DNA encoding-enhanced green fluorescent protein and then sonoporated in presence of contrast microbubbles. Twenty-four hours later, cells that received electrosonoporation demonstrated a four-fold increase in transfection level and a six-fold increase in transfection efficiency compared with cells having undergone electroporation alone. Although electroporation induced the formation of DNA aggregates into the cell membrane, sonoporation induced its direct propulsion into the cytoplasm. Sonoporation can improve the transfer of electro-induced DNA aggregates by allowing its free and rapid entrance into the cells. These results demonstrated that in vitro gene transfer by electrosonoporation could provide a new potent method for gene transfer.
Escoffre
J M
JM
CNRS, Institut de Pharmacologie et de Biologie Structurale, Toulouse, France.
Kaddur
K
K
Rols
M P
MP
Bouakaz
A
A
D002478:Cells, Cultured; D003287:Contrast Media / Q000008:administration & dosage; D018274:Electroporation / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D045423:Microbubbles; D010957:Plasmids / Q000235:genetics; D013010:Sonication / Q000379:methods*; D014161:Transduction, Genetic / Q000379:methods*; D014465:Ultrasonics / Q000379:methods*
D003287:Contrast Media; D049452:Green Fluorescent Proteins