Research Article | Published:

Gene transfer into muscle by electroporation in vivo

Nature Biotechnology volume 16, pages 867870 (1998) | Download Citation

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Abstract

Among the nonviral techniques for gene transfer in vivo, the direct injection of plasmid DNA into muscle is simple, inexpensive, and safe. Applications of this method have been limited by the relatively low expression levels of the transferred gene. We investigated the applicability of in vivo electroporation for gene transfer into muscle, using plasmid DNA expressing interleukin-5 (IL-5) as the vector. The tibialis anterior muscles of mice were injected with the plasmid DNA, and then a pair of electrode needles were inserted into the DNA injection site to deliver electric pulses. Five days later, the serum IL-5 levels were assayed. Mice that did not receive electroporation had serum levels of 0.2 ng/ml. Electroporation enhanced the levels to over 20 ng/ml. Histochemical analysis of muscles injected with a lacZ expression plasmid showed that in vivo electroporation increased both the number of muscle fibers taking up plasmid DNA and the copy number of plasmids introduced into the cells. These results demonstrate that gene transfer into muscle by electroporation in vivo is more efficient than simple intramuscular DNA injection.

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References

  1. 1.

    , , , , , et al. 1990. Direct gene transfer into mouse muscle in vivo. Science 247: 1465–1468.

  2. 2.

    , , , , and 1992. Long-term persistence of plasmid DNA and foreign gene expression in mouse muscle. Hum. Mol. Genet. 1: 363–369.

  3. 3.

    , , , , and 1993. Plasmid DNA is superior to viral vectors for direct gene transfer into adult mouse skeletal muscle. Hum. Gene Ther. 4: 733–740.

  4. 4.

    , , and 1993. Direct gene transfer into skeletal muscle in vivo: factors affecting efficiency of transfer and stability of expression. Hum. Gene Ther. 4: 151–159.

  5. 5.

    , , , , , et al. 1993. Heterologous protection against influenza by injection of DNA encoding a viral protein. Science 259: 1745–1749.

  6. 6.

    and 1995. Non-viral approaches to gene therapy. Br. Med. Bull. 51: 56–71.

  7. 7.

    , and 1995. Use of plasmid DNA for direct gene transfer and immunization. Ann. N.Y. Acad. Sci. 772: 21–29.

  8. 8.

    , , , , , and 1991. Conditions affecting direct gene transfer into rodent muscle in vivo. Biotechniques 11: 474–485.

  9. 9.

    and 1992. Age and sex influence expression of plasmid DNA directly injected into mouse skeletal muscle. FEBS Lett. 306: 203–205.

  10. 10.

    , , , , and 1994. Gene transfer in regenerating muscle. Hum. Gene Ther. 5: 11–18.

  11. 11.

    1993. Improved gene transfer by direct plasmid injection associated with regeneration in mouse skeletal muscle. FEBS Lett. 332: 179–182.

  12. 12.

    , , , , , et al. 1997. Intramuscular injection of expression plasmid DNA is an effective means of long-term systemic delivery of interleukin-5. Biochem. Biophys. Res. Commun. 233: 527–531.

  13. 13.

    , , , , , et al. 1996. Long-term expression of erythropoietin in the systemic circulation of mice after intramuscular injection of a plasmid DNA vector. Proc. Natl. Acad. Sci. USA 93: 10876–10880.

  14. 14.

    , , and 1991. In vivo electroporation and stable transformation of skin cells of newborn mice by plasmid DNA. Biochim. Biophys. Acta. 1088: 131–134.

  15. 15.

    , , , and 1997. Comparison of three nonviral transfection methods for foreign gene expression in early chicken embryos in ovo. Biochem. Biophys. Res. Commun. 230: 376–380.

  16. 16.

    , , , , , et al. 1996. In vivo electroinjection and expression in rat liver. FEBS Lett. 389: 225–228.

  17. 17.

    , , , , , and 1998. In vivo electrically mediated protein and gene transfer in murune melanoma. Nat. Biotechnol. 16: 168–171.

  18. 18.

    1992. Interleukin-5, eosinophils, and disease. Blood 79: 3101–3109.

  19. 19.

    1992. Interleukin-5. Curr. Opin. Immunol. 4: 299–306.

  20. 20.

    , , , , and 1997. “Green mice” as a source of ubiquitous green cells. FEBS Lett. 407: 313–319.

  21. 21.

    , , , , , et al. 1989. Expression vector system based on the chicken β-actin promoter directs efficient production of interleukin-5. Gene 79: 269–277.

  22. 22.

    , , and 1991. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108: 193–199.

  23. 23.

    , , , , and 1994. Control by pulse parameters of electric field-mediated gene transfer in mammalian cells. Biophys. J. 66: 524–531.

  24. 24.

    , , , , , et al. 1992. Direct gene transfer into nonhuman primate myofibers in vivo. Hum. Gene Ther. 3: 21–33.

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Author notes

    • Jun-ichi Miyazaki

    Corresponding author (e-mail: jimiyaza@nutri.med.osaka-u.ac.jp)

Affiliations

  1. Department of Nutrition and Physiological Chemistry, Osaka University Medical School, Suita, Osaka 565-0871, Japan.

    • Hiroyuki Aihara
    •  & Jun-ichi Miyazaki
  2. The Third Department of Internal Medicine, Tohoku University School of Medicine, Sendai, Miyagi 980-0872, Japan.

    • Hiroyuki Aihara

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DOI

https://doi.org/10.1038/nbt0998-867