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Epidermis-Targeted Gene Transfer Using In Vivo Electroporation

  • Hiroki Maruyama
  • Jun-Ichi Miyazaki
  • Fumitake Gejyo
Part of the Methods in Molecular Biology™ book series (MIMB, volume 289)

Abstract

The skin is an important target for gene transfer because of its easy accessibility. Recently, we demonstrated that an in vivo electroporation approach could be applied to plasmid deoxyribonucleic acid delivery in rat skin of the abdominal area. Plate-and-fork type electrodes were effective for gene delivery by skin-targeted gene transfer. Eight 12–24 V, 50-m electric pulses were delivered through the electrodes. Skin injection with pCAGGS-lacZ showed lacZ gene expression in the upper most cell layers (horny, granular, and prickle cell layers) of the epidermis on day 1 and in the subcutaneous muscle layer on day 7 after the pCAGGS-lacZ transfer. Slight skin damage as the result of the gene transfer procedure was evident on day 1 but absent by day 7. These results demonstrate that plasmid deoxyribonucleic acid transfer by in vivo electroporation at low voltage is a useful procedure for short-term skin-targeted gene transfer.

Key Words

Electroporation gene transfer skin plasmid DNA epidermis electrodes CAG promoter 

References

  1. 1.
    Hengge, U. R, Chan E. F., Foster, R. A., Walker, P. S., and Vogel, J. C. (1995) Cytokine gene expression in epidermis with biological effects following injection of naked DNA. Nat. Genet. 10, 161–166.PubMedCrossRefGoogle Scholar
  2. 2.
    Hengge, U. R., Walker, P. S., and Vogel, J. C. (1996) Expression of naked DNA in human, pig, and mouse skin. J. Clin. Invest. 97, 2911–2916.PubMedCrossRefGoogle Scholar
  3. 3.
    Li, L. and Hoffmann, R. M. (1995) The feasibility of targeted selective gene therapy of the hair follicle. Nat. Med. 1, 705–706.PubMedCrossRefGoogle Scholar
  4. 4.
    Vogel, J. C. (1999) A direct in vivo approach for skin gene therapy. P. Assoc. Am. Phys. 111, 190–197.CrossRefGoogle Scholar
  5. 5.
    Wolff, J. A., Malone, R. W., Williams, P., Chong, W., Acsadi, G., Jani, A., et al. (1990) Direct gene transfer into mouse muscle in vivo. Science 247, 1465–1468.PubMedCrossRefGoogle Scholar
  6. 6.
    Raz, E., Carson, D. A., Parker, S. E., Parr, T. B., Abai, A. M., Aichinger, G, et al. (1994) Intradermal gene immunization: the possible role of DNA uptake in the induction of cellular immunity to viruses. Proc. Natl. Acad. Sci. USA 91, 9519–9523.PubMedCrossRefGoogle Scholar
  7. 7.
    Fan, H., Lin, Q., Morrissey, G R., and Khavari, P. A. (1998) Immunization via hair follicles by topical application of naked DNA to normal skin. Nat. Biotechnol. 17, 870–872.Google Scholar
  8. 8.
    Maruyama, H., Ataka, K., Higuchi, N, Sakamoto, F, Gejyo, F, and Miyazaki, J. (2001) Skin-targeted gene transfer using in-vivo electroporation. Gene Ther. 8, 1808–1812.PubMedCrossRefGoogle Scholar
  9. 9.
    Niwa, H., Yamamura, K., and Miyazaki, J. (1991) Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108, 193–199.PubMedCrossRefGoogle Scholar
  10. 10.
    Aihara, H. and Miyazaki, J. (1998) Gene transfer into muscle by electroporation in vivo. Nat. Biotechnol. 16, 867–870.PubMedCrossRefGoogle Scholar
  11. 11.
    Maruyama, H., Higuchi, N, Nishikawa, U., Hirahara, H., Iino, N, Kameda, S., et al. (2002) Kidney-targeted naked DNA transfer by retrograde renal vein injection in rats. Hum. Gene Ther. 13, 455–468.PubMedCrossRefGoogle Scholar
  12. 12.
    Sambrook, J. and Russell, D. W. (2001) β-galactosidase, in Molecular Cloning: A Laboratory Manual, 3rd ed., vol. 3 (Argentine, J., ed.), Cold Spring Harbor Laboratory Press, New York, pp. 17.97–17.99.Google Scholar

Copyright information

© Humana Press Inc. 2005

Authors and Affiliations

  • Hiroki Maruyama
    • 1
  • Jun-Ichi Miyazaki
    • 2
  • Fumitake Gejyo
    • 1
  1. 1.Division of Clinical Nephrology and RheumatologyNiigata University Graduate School of Medical and Dental SciencesNiigataJapan
  2. 2.Division of Stem Cell Regulation ResearchOsaka University Medical SchoolOsakaJapan

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