Abstract
Electroporation is an effective physical delivery method. A variety of factors have been shown to affect the electroporation-mediated gene delivery efficiency. Here we report the usefulness of noncoding short-fragment DNA (sf-DNA) for facilitating electroporation-mediated gene transfer. The plasmid pGL3-control encoding firefly luciferase was injected into tissue together with or without sf-DNA in different length or dose. Immediately after injection, the tissues were electroporated and the level of luciferase activity was assessed 24 h later. The results showed that plasmid DNA formulated with sf-DNA resulted in significant improvement in electroporation-mediated gene transfer efficiency. The effect is dose and length dependent, and also found in low-voltage electroporation. These results indicated that sf-DNA can be used as a helper molecule to improve the electroporation-mediated gene transfection efficiency.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Sukharev SI, Klenchin VA, Serov SM et al (1992) Electroporation and electrophoretic DNA transfer into cells. The effect of DNA interaction with electropores. Biophys J 63:1320–1327
Titomirov AV, Sukharev S, Kistanova E (1991) In vivo electroporation and stable transformation of skin cells of newborn mice by plasmid DNA. Biochim Biophys Acta 1088:131–134
Zhao X (2000) Gene transfer and drug delivery by electronic pulse delivery. A nonviral delivery approach. Methods Mol Biol 133:37–43
Lladser A, Ljungberg K, Tufvesson H et al (2010) Intradermal DNA electroporation induces survivin-specific CTLs, suppresses angiogenesis and confers protection against mouse melanoma. Cancer Immunol Immunother 59:81–92
Yin J, Dai A, Lecureux J et al (2010) High antibody and cellular responses induced to HIV-1 clade C envelope following DNA vaccines delivered by electroporation. Vaccine 29:6763–6770
Ferraro B, Cruz YL, Coppola D et al (2009) Intradermal delivery of plasmid VEGF(165) by electroporation promotes wound healing. Mol Ther 17:651–657
Daud AI, DeConti RC, Andrews S et al (2008) Phase I trial of interleukin-12 plasmid electroporation in patients with metastatic melanoma. J Clin Oncol 26:5896–5903
Chiarella P, Fazio VM, Signori E (2010) Application of electroporation in DNA vaccination protocols. Curr Gene Ther 10:281–286
Gehl J, Mir LM (1999) Determination of optimal parameters for in vivo gene transfer by electroporation, using a rapid in vivo test for cell permeabilization. Biochem Biophys Res Commun 261:377–380
Mir LM, Bureau MF, Rangara R et al (1998) Long-term, high level in vivo gene expression after electric pulse-mediated gene transfer into skeletal muscle. C R Acad Sci III 321:893–899
Lucas ML, Jaroszeski MJ, Gilbert R et al (2001) In vivo electroporation using an exponentially enhanced pulse: a new waveform. DNA Cell Biol 20:183–188
Zhang L, Nolan E, Kreitschitz S et al (2002) Enhanced delivery of naked DNA to the skin by non-invasive in vivo electroporation. Biochim Biophys Acta 1572:1–9
Broderick KE, Shen X, Soderholm J et al (2011) Prototype development and preclinical immunogenicity analysis of a novel minimally invasive electroporation device. Gene Ther 18:258–265
Lee MJ, Cho SS, Jang HS et al (2002) Optimal salt concentration of vehicle for plasmid DNA enhances gene transfer mediated by electroporation. Exp Mol Med 34:265–272
Chesnoy S, Huang L (2002) Enhanced cutaneous gene delivery following intradermal injection of naked DNA in a high ionic strength solution. Mol Ther 5:57–62
McMahon JM, Signori E, Wells KE et al (2001) Optimisation of electrotransfer of plasmid into skeletal muscle by pretreatment with hyaluronidase – increased expression with reduced muscle damage. Gene Ther 8:1264–1270
Sen A, Zhao Y, Zhang L et al (2002) Enhanced transdermal transport by electroporation using anionic lipids. J Control Release 82:399–405
Nicol F, Wong M, MacLaughlin FC et al (2002) Poly-L-glutamate, an anionic polymer, enhances transgene expression for plasmids delivered by intramuscular injection with in vivo electroporation. Gene Ther 9:1351–1358
Peng J, Zhao Y, Mai J et al (2012) Short noncoding DNA fragment improve efficiencies of in vivo electroporation-mediated gene transfer. J Gene Med 14:563–569
Acknowledgment
This work was supported by Chinese National Natural Science Foundation (Grant No. 30801446).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Peng, J., Zhao, Y., Xu, Y. (2014). Short-Fragment DNA-Mediated In Vivo DNA Electroporation Delivery. 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_5
Download citation
DOI: https://doi.org/10.1007/978-1-4614-9632-8_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4614-9631-1
Online ISBN: 978-1-4614-9632-8
eBook Packages: Springer Protocols