{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T00:55:20Z","timestamp":1770339320685,"version":"3.49.0"},"reference-count":30,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2020,9,5]],"date-time":"2020-09-05T00:00:00Z","timestamp":1599264000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"POR Campania FESR 2014 2020","award":["CUP: B63D18000560007"],"award-info":[{"award-number":["CUP: B63D18000560007"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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.<\/jats:p>","DOI":"10.3390\/ijms21186494","type":"journal-article","created":{"date-parts":[[2020,9,6]],"date-time":"2020-09-06T23:12:49Z","timestamp":1599433969000},"page":"6494","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Definition of a Novel Plasmid-Based Gene Transfection Protocol of Mammalian Skeletal Muscles by Means of In Vivo Electroporation"],"prefix":"10.3390","volume":"21","author":[{"given":"Enrico P.","family":"Spugnini","sequence":"first","affiliation":[{"name":"Biopulse srl, 80100 Naples, Italy"},{"name":"Orchidealab srl, 00100 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0585-1309","authenticated-orcid":false,"given":"Manuel","family":"Scimeca","sequence":"additional","affiliation":[{"name":"Department of Biomedicine and Prevention, University of Rome \u201cTor Vergata\u201d 00100 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bruno","family":"Amadio","sequence":"additional","affiliation":[{"name":"UOSD SAFU Department of Research, Diagnosis and Innovative Technologies, Translational Research Area, IRCCS Regina Elena National Cancer Institute, 00100 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Giancarlo","family":"Cortese","sequence":"additional","affiliation":[{"name":"UOSD SAFU Department of Research, Diagnosis and Innovative Technologies, Translational Research Area, IRCCS Regina Elena National Cancer Institute, 00100 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5281-4191","authenticated-orcid":false,"given":"Maurizio","family":"Fanciulli","sequence":"additional","affiliation":[{"name":"UOSD SAFU Department of Research, Diagnosis and Innovative Technologies, Translational Research Area, IRCCS Regina Elena National Cancer Institute, 00100 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bruno","family":"Vincenzi","sequence":"additional","affiliation":[{"name":"Department of Medical Oncology, Campus Bio-Medico University, 00100 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3905-6154","authenticated-orcid":false,"given":"Antonio","family":"De Luca","sequence":"additional","affiliation":[{"name":"Department of Mental and Physical Health and Preventive Medicine, Section of Human Anatomy, University of Campania \u201cL. Vanvitelli\u201d, 80100 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8693-3842","authenticated-orcid":false,"given":"Alfonso","family":"Baldi","sequence":"additional","affiliation":[{"name":"Biopulse srl, 80100 Naples, Italy"},{"name":"Orchidealab srl, 00100 Rome, Italy"},{"name":"Department of Environmental, Biological, Pharmaceutical Sciences and Technologies, University of Campania \u201cL. 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Develop."},{"key":"ref_4","first-page":"491","article-title":"Viral vectors for gene transfer to striated muscle","volume":"6","author":"Gregoveric","year":"2004","journal-title":"Curr. Opin. Mol. Ther."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1016\/j.ymthe.2005.09.019","article-title":"Optimizing plasmid-based gene transfer for investigating skeletal muscle structure and function","volume":"13","author":"Schertzer","year":"2005","journal-title":"Mol. Ther."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"155","DOI":"10.2165\/00063030-200418030-00002","article-title":"Electroporation for gene transfer to skeletal muscles: Current status","volume":"18","author":"McMahon","year":"2004","journal-title":"BioDrugs"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"105204","DOI":"10.1016\/j.cmpb.2019.105204","article-title":"Numerical optimization of plasmid DNA delivery combined with hyaluronidase injection for electroporation protocol","volume":"186","author":"Peri","year":"2020","journal-title":"Comput. Methods Programs Biomed."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1007\/978-1-59745-237-3_7","article-title":"Plasmid-based gene transfer in mouse skeletal muscle by electroporation","volume":"433","author":"Schertzer","year":"2008","journal-title":"Methods Mol. Biol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1619","DOI":"10.1038\/sj.gt.3302827","article-title":"Variability of naked DNA expression after direct local injection: The influence of the injection speed","volume":"13","author":"Andre","year":"2006","journal-title":"Gene Ther."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1171","DOI":"10.1038\/sj.gt.3301226","article-title":"Sodium phosphate enhances plasmid DNA expression in vivo","volume":"7","author":"Hartikka","year":"2000","journal-title":"Gene Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2097","DOI":"10.1096\/fj.07-093765","article-title":"Nuclear-targeted chimeric vector enhancing nonviral gene transfer into skeletal muscle of Fabry mice in vivo","volume":"22","author":"Lavigne","year":"2008","journal-title":"FASEB J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1016\/j.ymthe.2004.06.642","article-title":"Factors influencing the efficacy, longevity, and safety of electroporation-assisted plasmid-based gene transfer into mouse muscles","volume":"10","author":"Molnar","year":"2004","journal-title":"Mol. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2133","DOI":"10.1038\/sj.gt.3302125","article-title":"Enhanced systemic transgene expression after nonviral salivary gland transfection using a novel endonuclease inhibitor\/DNA formulation","volume":"10","author":"Niedzinski","year":"2003","journal-title":"Gene Ther."},{"key":"ref_14","first-page":"1520","article-title":"DNA transfection of mammalian skeletal muscles using in vivo electroporation","volume":"32","author":"DiFranco","year":"2009","journal-title":"J. Vis. Exp."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"21","DOI":"10.5826\/dpc.0703a04","article-title":"Successful treatment of plantar warts with intralesional bleomycin and electroporation: Pilot prospective study","volume":"7","author":"Pasquali","year":"2017","journal-title":"Dermatol. Pract. Concept"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1007\/s13555-018-0222-9","article-title":"Successful Treatment of a Keratoacanthoma with Electrochemotherapy: A Case Report","volume":"8","author":"Pasquali","year":"2018","journal-title":"Dermatol. Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1002\/jcp.25505","article-title":"Novel Instruments for the Implementation of Electrochemotherapy Protocols: From Bench Side to Veterinary Clinic","volume":"232","author":"Spugnini","year":"2017","journal-title":"J. Cell Physiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1016\/j.cvsm.2019.04.006","article-title":"Electrochemotherapy in Veterinary Oncology: State-of-the-Art and Perspectives","volume":"49","author":"Spugnini","year":"2019","journal-title":"Vet. Clin. North Am. Small Animal Pract."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.1016\/j.joca.2016.03.008","article-title":"Gait analysis methods for rodent models of arthritic disorders: Reviews and recommendations","volume":"24","author":"Lakes","year":"2016","journal-title":"Osteoarthr. Cartil."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Hobernik, D., and Bros, M. (2018). DNA Vaccines\u2014How Far From Clinical Use?. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19113605"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.7774\/cevr.2015.4.1.1","article-title":"Improvement of DNA vaccination by adjuvants and sophisticated delivery devices: Vaccine-platforms for the battle against infectious diseases","volume":"4","author":"Grunwald","year":"2015","journal-title":"Clin. Exp. Vaccine Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"C1239","DOI":"10.1152\/ajpcell.00431.2010","article-title":"Physiological and histological changes in skeletal muscle following in vivo gene transfer by electroporation","volume":"301","author":"Roche","year":"2011","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Sokolowska, E., and Blachnio-Zabielska, A.U. (2019). A critical review of electroporation as a plasmid delivery system in mouse skeletal muscle. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20112776"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"BSR20160616","DOI":"10.1042\/BSR20160616","article-title":"Cytoplasmic transport and nuclear import of plasmid DNA","volume":"37","author":"Bai","year":"2017","journal-title":"Biosci. Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3446","DOI":"10.1002\/jcb.24221","article-title":"Role of FAP48 in HIV-associated lipodystrophy","volume":"113","author":"Esposito","year":"2012","journal-title":"J. Cell Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.vascn.2016.10.005","article-title":"Using Tiletamine-Zolazepam-Xylazine Anesthesia Compared to CO2-inhalation for Terminal Clinical Chemistry, Hematology, and Coagulation Analysis in Mice","volume":"84","author":"Khokhlova","year":"2017","journal-title":"J. Pharmacol. Toxicol. Methods"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1016\/j.cbi.2019.03.031","article-title":"Effects of curcumin and its adjuvant on TPC1 thyroid cell line","volume":"305","author":"Esposito","year":"2019","journal-title":"Chem. Biol. Interact."},{"key":"ref_28","first-page":"217","article-title":"Small leucine rich proteoglycans are differently distributed in normal and pathological endometrium","volume":"29","author":"Lucariello","year":"2015","journal-title":"In Vivo"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"885","DOI":"10.1369\/jhc.3A6186.2004","article-title":"The serine protease HtrA1 is upregulated in the human placenta during pregnancy","volume":"52","author":"Fedele","year":"2004","journal-title":"J. Histochem. Cytochem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3175","DOI":"10.1158\/1078-0432.CCR-04-0631","article-title":"Ferritin contributes to melanoma progression by modulating cell growth and sensitivity to oxidative stress","volume":"11","author":"Baldi","year":"2005","journal-title":"Clin. 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