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The Journal of Membrane Biology

, Volume 248, Issue 5, pp 919–928 | Cite as

Improved Specificity of Gene Electrotransfer to Skin Using pDNA Under the Control of Collagen Tissue-Specific Promoter

  • Spela Kos
  • Natasa Tesic
  • Urska Kamensek
  • Tanja Blagus
  • Maja Cemazar
  • Simona Kranjc
  • Jaka Lavrencak
  • Gregor SersaEmail author
Article

Abstract

In order to ensure safe, efficient and controlled gene delivery to skin, the improvement of delivery methods together with proper design of DNA is required. Non-viral delivery methods, such as gene electrotransfer, and the design of tissue-specific promoters are promising tools to ensure the safety of gene delivery to the skin. In the scope of our study, we evaluated a novel skin-specific plasmid DNA with collagen (COL) promoter, delivered to skin cells and skin tissue by gene electrotransfer. In vitro, we determined the specificity of the COL promoter in fibroblast cells. The specific expression under the control of COL promoter was obtained for the reporter gene DsRed as well as for therapeutic gene encoding cytokine IL-12. In vivo, the plasmid with COL promoter encoding the reporter gene DsRed was efficiently transfected to mouse skin. It resulted in the notable and controlled manner, however, in lower and shorter expression, compared to that obtained with ubiquitous promoter. The concentration of the IL-12 in the skin after the in vivo transfection of plasmid with COL promoter was in the same range as after the treatment in control conditions (injection of distilled water followed by the application of electric pulses). Furthermore, this gene delivery was local, restricted to the skin, without any evident systemic shedding of IL-12. Such specific targeting of skin cells, observed with tissue-specific COL promoter, would improve the effectiveness and safety of cutaneous gene therapies and DNA vaccines.

Keywords

Skin gene electrotransfer Skin-specific promoter Collagen promoter Interleukin-12 Electroporation Gene delivery 

Notes

Acknowledgments

The authors acknowledge the financial support from the state budget by the Slovenian Research Agency (Program No. P3-0003, Projects Nos. J3-4211, J3-6793, J3-4259, J3-6796). The research was conducted in the scope of LEA EBAM (French-Slovenian European Associated Laboratory: Pulsed Electric Fields Applications in Biology and Medicine) and is a result of networking efforts within COST TD1104 Action. We would like to thank Mira Lavric (Institute of Oncology Ljubljana, Ljubljana, Slovenia) for all the valuable work she contributed to this research.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Spela Kos
    • 1
  • Natasa Tesic
    • 2
  • Urska Kamensek
    • 1
  • Tanja Blagus
    • 1
  • Maja Cemazar
    • 1
    • 2
  • Simona Kranjc
    • 1
  • Jaka Lavrencak
    • 3
  • Gregor Sersa
    • 1
    Email author
  1. 1.Department of Experimental OncologyInstitute of Oncology LjubljanaLjubljanaSlovenia
  2. 2.Faculty of Health SciencesUniversity of PrimorskaIzolaSlovenia
  3. 3.Department of CytopathologyInstitute of Oncology LjubljanaLjubljanaSlovenia

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