Long-term, high level in vivo gene expression after electric pulse-mediated gene transfer into skeletal muscle.

Gene delivery to skeletal muscle is a promising strategy for the treatment of muscle disorders and for the local or systemic secretion of therapeutic proteins. However, current DNA delivery technologies have to be improved. We report very efficient luciferase gene transfer into muscle fibres obtained through the delivery of square-wave electric pulses of moderate field strength (100-200 V/cm) and of long duration (20 ms) to muscle previously injected with plasmid DNA. This intramuscular 'electrotransfer' method increases reporter gene expression by more than 100 times. It is noteworthy that this expression remains high and stable for at least 9 months. Moreover, intramuscular electrotransfer strongly decreases the interindividual variability usually observed after plasmid DNA injection into muscle fibres. Therefore, DNA electrotransfer in muscle possesses broad potential applications in gene therapy and for physiological, pharmacological and developmental studies.

[1]  J. Gehl,et al.  Enhancement of cytotoxicity by electropermeabilization: an improved method for screening drugs , 1998, Anti-cancer drugs.

[2]  Z. Bebők,et al.  Cationic lipids for reporter gene and CFTR transfer to rat pulmonary epithelium. , 1995, Gene therapy.

[3]  H. Aihara,et al.  Gene transfer into muscle by electroporation in vivo , 1998, Nature Biotechnology.

[4]  E. Svensson,et al.  Long-term expression of erythropoietin in the systemic circulation of mice after intramuscular injection of a plasmid DNA vector. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Yilin Zhang,et al.  Toward development of a non-viral gene therapeutic. , 1997, Advanced drug delivery reviews.

[6]  Christian Domenge,et al.  L'électrochimiothérapie, un nouveau traitement antitumoral : premier essai clinique , 1991 .

[7]  G. Acsadi,et al.  Direct gene transfer into mouse muscle in vivo. , 1990, Science.

[8]  S. Sukharev,et al.  In vivo electroporation and stable transformation of skin cells of newborn mice by plasmid DNA. , 1991, Biochimica et biophysica acta.

[9]  D Miklavcic,et al.  Electrochemotherapy with cisplatin: potentiation of local cisplatin antitumour effectiveness by application of electric pulses in cancer patients. , 1998, European journal of cancer.

[10]  Leaf Huang,et al.  Lipidic vector systems for gene transfer. , 1997, Critical reviews in therapeutic drug carrier systems.

[11]  A Brun,et al.  A new brain tumour therapy combining bleomycin with in vivo electropermeabilization. , 1993, Biochemical and biophysical research communications.

[12]  Darius Moradpour,et al.  In vivo gene electroinjection and expression in rat liver , 1996, FEBS letters.

[13]  Toshiyuki Matsuzaki,et al.  Direct gene transfer into rat liver cells by in vivo electroporation , 1998, FEBS letters.

[14]  J. Northrop,et al.  Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[15]  L. Chernomordik,et al.  Electroporation and electrophoretic DNA transfer into cells. The effect of DNA interaction with electropores. , 1992, Biophysical journal.

[16]  G. Repetto,et al.  Expression of naked plasmids by cultured myotubes and entry of plasmids into T tubules and caveolae of mammalian skeletal muscle. , 1992, Journal of cell science.

[17]  D. Miklavcic,et al.  Effective treatment of cutaneous and subcutaneous malignant tumours by electrochemotherapy. , 1998, British Journal of Cancer.

[18]  E. Neumann,et al.  Gene transfer into mouse lyoma cells by electroporation in high electric fields. , 1982, The EMBO journal.

[19]  L. Mir,et al.  Cell electropermeabilization: a new tool for biochemical and pharmacological studies. , 1993, Biochimica et biophysica acta.

[20]  M J Jaroszeski,et al.  Novel electrode designs for electrochemotherapy. , 1997, Biochimica et biophysica acta.

[21]  P. Briand,et al.  Efficient adenovirus-mediated transfer of a human minidystrophin gene to skeletal muscle of mdx mice , 1993, Nature.

[22]  H. Potter,et al.  Electroporation in biology: methods, applications, and instrumentation. , 1988, Analytical biochemistry.

[23]  M. Jaroszeski,et al.  Treatment of cutaneous and subcutaneous tumors with electrochemotherapy using intralesional bleomycin , 1998, Cancer.

[24]  E. Nabel,et al.  Site-specific gene expression in vivo by direct gene transfer into the arterial wall. , 1990, Science.

[25]  M. Rols,et al.  In vivo electrically mediated protein and gene transfer in murine melanoma , 1998, Nature Biotechnology.