TY - JOUR
T1 - Definition of a novel plasmid-based gene transfection protocol of mammalian skeletal muscles by means of in vivo electroporation
AU - Spugnini, Enrico P.
AU - Scimeca, Manuel
AU - Amadio, Bruno
AU - Cortese, Giancarlo
AU - Fanciulli, Maurizio
AU - Vincenzi, Bruno
AU - De Luca, Antonio
AU - Baldi, Alfonso
N1 - Funding Information:
Funding: This work was partially supported by POR Campania FESR 2014 2020 CUP: B63D18000560007.
Publisher Copyright:
© 2020 by the authors. Licensee MDPI, Basel, Switzerland.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/9/2
Y1 - 2020/9/2
N2 - 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.
AB - 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.
KW - Biphasic pulses
KW - DNA plasmid
KW - Electroporation
KW - Green Fluorescent Protein (GFP)
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U2 - 10.3390/ijms21186494
DO - 10.3390/ijms21186494
M3 - Article
C2 - 32899477
AN - SCOPUS:85090200540
VL - 21
SP - 1
EP - 11
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
SN - 1661-6596
IS - 18
M1 - 6494
ER -