Definition of a novel plasmid-based gene transfection protocol of mammalian skeletal muscles by means of in vivo electroporation

Enrico P. Spugnini, Manuel Scimeca, Bruno Amadio, Giancarlo Cortese, Maurizio Fanciulli, Bruno Vincenzi, Antonio De Luca, Alfonso Baldi

Research output: Contribution to journalArticlepeer-review

Abstract

We describe an original electroporation protocol for in vivo plasmid DNA transfection. The right hind limbs of C57 mice are exposed to a specifically designed train of permeabilizing electric pulses by transcutaneous application of tailored needle electrodes, immediately after the injection of pEGFP-C1 plasmid encoding GFP (Green Fluorescente Protein). The electroporated rodents show a greater GFP expression than the controls at three different time points (4, 10, and 15 days). The electroporated muscles display only mild interstitial myositis, with a significant increase in inflammatory cell infiltrates. Finally, mild gait abnormalities are registered in electroporated mice only in the first 48 h after the treatment. This protocol has proven to be highly efficient in terms of expression levels of the construct, is easy to apply since it does not require surgical exposure of the muscle and is well tolerated by the animals because it does not cause evident morphological and functional damage to the electroporated muscle.

Original languageEnglish
Article number6494
Pages (from-to)1-11
Number of pages11
JournalInternational Journal of Molecular Sciences
Volume21
Issue number18
DOIs
Publication statusPublished - Sep 2 2020

Keywords

  • Biphasic pulses
  • DNA plasmid
  • Electroporation
  • Green Fluorescent Protein (GFP)

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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