Abstract
Electroporation is a safe, efficient, and inexpensive method to transfer naked plasmid DNA into various tissues. For electroporation-mediated gene transfer to the mouse lung, a plasmid solution is delivered to the lungs via the trachea. Immediately after plasmid delivery, eight square wave pulses are delivered by two pre-gelled electrodes placed on each side of the chest. An optimal field strength in mice is 200 V/cm, with a pulse duration of 10 ms each and a 1 s interval between pulses. High level gene expression can be achieved within 24 h in all cell types in the lung with very little inflammation and no apparent trauma.
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References
Somiari S, Glasspool-Malone J, Drabick JJ et al (2000) Theory and in vivo application of electroporative gene delivery. Mol Ther 2: 178–187
Heller L, Jaroszeski MJ, Coppola D, Pottinger C, Gilbert R, Heller R (2000) Electrically mediated plasmid DNA delivery to hepatocellular carcinomas in vivo. Gene Ther 7:826–829
Cemazar M, Golzio M, Sersa G et al (2009) Control by pulse parameters of DNA electrotransfer into solid tumors in mice. Gene Ther 16:635–644
Wells JM, Li LH, Sen A, Jahreis GP, Hui SW (2000) Electroporation-enhanced gene delivery in mammary tumors. Gene Ther 7: 541–547
Lin F, Shen X, Kichaev G et al (2012) Optimization of electroporation-enhanced intradermal delivery of DNA vaccine using a minimally invasive surface device. Hum Gene Ther Methods 23:157–168
Dean DA (2003) Electroporation of the vasculature and the lung. DNA Cell Biol 22:797–806
West J, Rodman DM (2001) Gene therapy for pulmonary diseases. Chest 119:613–617
Weiss D (2002) Delivery of gene transfer vectors to lung: obstacles and the role of adjunct techniques for airway administration. Mol Ther 6:148–152
Dean DA, Machado-Aranda D, Blair-Parks K, Yeldandi AV, Young JL (2003) Electroporation as a method for high-level non-viral gene transfer to the lung. Gene Ther 10:1608–1615
Machado-Aranda D, Adir Y, Young JL et al (2005) Gene transfer of the na+, k+-atpase b1 subunit using electroporation increases lung liquid clearance in rats. Am J Respir Crit Care Med 171:204–211
Jones MR, Simms BT, Lupa MM, Kogan MS, Mizgerd JP (2005) Lung nf-kappab activation and neutrophil recruitment require il-1 and tnf receptor signaling during pneumococcal pneumonia. J Immunol 175:7530–7535
Gazdhar A, Bilici M, Pierog J et al (2006) In vivo electroporation and ubiquitin promoter–a protocol for sustained gene expression in the lung. J Gene Med 8:910–918
Mutlu GM, Machado-Aranda D, Norton JE et al (2007) Electroporation-mediated gene transfer of the na+, k+-atpase rescues endotoxin-induced lung injury. Am J Respir Crit Care Med 176:582–590
Zhou R, Norton JE, Zhang N, Dean DA (2007) Electroporation-mediated transfer of plasmids to the lung results in reduced tlr9 signaling and inflammation. Gene Ther 14: 775–780
Pringle IA, McLachlan G, Collie DD et al (2007) Electroporation enhances reporter gene expression following delivery of naked plasmid DNA to the lung. J Gene Med 9: 369–380
Gazdhar A, Fachinger P, van Leer C et al (2007) Gene transfer of hepatocyte growth factor by electroporation reduces bleomycin-induced lung fibrosis. Am J Physiol Lung Cell Mol Physiol 292:L529–L536
Gazdhar A, Temuri A, Knudsen L et al (2013) Targeted gene transfer of hepatocyte growth factor to alveolar type ii epithelial cells reduces lung fibrosis in rats. Hum Gene Ther 24:105–116
Leuenberger A, Gazdhar A, Herrmann G, Ochs M, Geiser T, Knudsen L (2012) Cell-specific expression of human hgf by alveolar type ii cells induces remodeling of septal wall tissue in the lung: a morphometric study. J Appl Physiol 113:799–807
O’Reilly MA, Yee M, Buczynski BW et al (2012) Neonatal oxygen increases sensitivity to influenza a virus infection in adult mice by suppressing epithelial expression of ear1. Am J Pathol 181:441–451
Degiulio JV, Kaufman CD, Dean DA (2010) The sp-c promoter facilitates alveolar type ii epithelial cell-specific plasmid nuclear import and gene expression. Gene Ther 17: 541–549
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Young, J.L., Barravecchia, M.S., Dean, D.A. (2014). Electroporation-Mediated Gene Delivery to the Lungs. In: Li, S., Cutrera, J., Heller, R., Teissie, J. (eds) Electroporation Protocols. Methods in Molecular Biology, vol 1121. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4614-9632-8_17
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DOI: https://doi.org/10.1007/978-1-4614-9632-8_17
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4614-9631-1
Online ISBN: 978-1-4614-9632-8
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