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Epidermis-targeted gene transfer using in vivo electroporation
Hiroki Maruyama1, Jun-Ichi Miyazaki, Fumitake Gejyo
1Division of Clinical Nephrology and Rheumatology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
Methods in Molecular Biology (Clifton, N.J.)
|October 27, 2004
Summary
This study shows low-voltage in vivo electroporation effectively delivers plasmid deoxyribonucleic acid (DNA) to rat skin. This method enables short-term gene expression in epidermal and muscle layers with minimal transient skin damage.
Area of Science:
- Dermatology
- Molecular Biology
- Biotechnology
Background:
- The skin's accessibility makes it a prime target for gene transfer applications.
- Previous methods for gene delivery to the skin have limitations.
- In vivo electroporation offers a potential non-viral gene delivery strategy.
Purpose of the Study:
- To evaluate the efficacy of in vivo electroporation for plasmid deoxyribonucleic acid (DNA) delivery in rat skin.
- To assess gene expression patterns and potential side effects of the procedure.
Main Methods:
- Plasmid DNA (pCAGGS-lacZ) was delivered to rat abdominal skin using plate-and-fork electrodes.
- In vivo electroporation was performed with eight 12-24 V, 50-ms electric pulses.
- Gene expression was analyzed on days 1 and 7 post-transfer.
Main Results:
- LacZ gene expression was observed in the epidermis on day 1 and in the subcutaneous muscle on day 7.
- The gene transfer procedure caused slight, transient skin damage that resolved by day 7.
- Low-voltage electroporation facilitated effective plasmid DNA delivery.
Conclusions:
- In vivo electroporation is a viable technique for short-term, skin-targeted gene transfer.
- This method demonstrates efficient delivery of plasmid DNA to epidermal and muscle layers.
- The procedure is well-tolerated with minimal and temporary side effects.