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http://dx.doi.org/10.4333/KPS.2004.34.4.263

Cellular Uptake and Transfection Efficiency of Plasmid DNA Using Low Molecular Weight Polyethylenimine  

Jeong, Gil-Jae (College of Medicine, Pochon CHA University)
Park, Kui-Lye (National Institute of Toxicological Research)
Shin, Ji-Young (College of Medicine, Pochon CHA University)
Choi, Han-Gon (College of Pharmacy, Yeungnam University)
Oh, Yu-Kyoung (College of Medicine, Pochon CHA University)
Publication Information
Journal of Pharmaceutical Investigation / v.34, no.4, 2004 , pp. 263-267 More about this Journal
Abstract
Branched and linear polyethylenimines (PEIs) have been studied as efficient and versatile agents for gene delivery in vitro and in vivo. PEIs exist in a linear or branched topology and are available in a wide range of molecular weight (Mw). Most studies have been done using PEIs with Mw higher than 10Kd. This study was aimed to test the transfection efficiency and the cell viability following gene delivery using PEI of Mw 2Kd, a relatively lower Mw cationic polymer. We used murine interleukin-2(mIL-2) plasmid DNA complexed with branched PEI 2Kd or 25Kd, and transfected them into a myoblast muscle cell line, C2C12. The cellular uptake of mIL-2 plasmid DNA was determined using quantitative polymerase chain reaction. RNA transcript levels were studied in the myoblast cells. Our results show that PEI 2Kd was as effective as PEI 25Kd in celluar gene delivery and transfection efficiency in C2C12 cells. Moreover, MTT assay indicated that PEI 2Kd/DNA complexes did not significantly reduce the cell viability regardless of N/P ratios. These results suggest that PEI of Mw 2Kd might play a role as effective and low toxic nonviral vector systems for muscular cell lines.
Keywords
Polyethylenimine; Low molecular weight polymer; Cellular uptake; Transfection; Cell viability;
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1 B. Pitard, H. Pollard, O. Agbulut, O. Lambert, J.T. Vilquin, Y Cherel, J. Abadie, J.L. Samuel, J.L. Rigaud, S. Menoret, I. Anegon and D.A Escande, Nonionic amphiphile agent promotes gene delivery. in vivo to skeletal and cardiac muscles, Hum. Gene Ther., 13, 1767-1775 (2002)   DOI   ScienceOn
2 H. Pollard, J.S. Remy, G. Loussouam, S. Demolombe, J.P Behr and D. Escande, Polyethylenimine but not cationic lipids promotes transgene delivery to the nucleus in mammalian cells, BioI. Chem. J., 273, 7507-7511 (1998)   DOI   ScienceOn
3 WT. Godbey, K.K Wu and A.G. Mikos, Poly(ethylenimine)-mediated gene delivery affects endothelial cell function and viability, Biomaterials, 22, 471-480 (2001)   DOI   ScienceOn
4 C.A. Josephine, S. Brian and L. Jack, Muskelin, a novel intracellular mediator of cell adhesive and cytoskeletal responses to thrombospondin-1, EMBO J., 17, 4964-4974 (1998)   DOI   ScienceOn
5 D.A Treeo and RF Selden, Non-viral gene therapy, Mol. Med. Today, 1, 314-321 (1995)   DOI   ScienceOn
6 R.S. Kevin, Gene therapy: theoretical and bioethical concepts, Gene Ther., 34, 247-268 (2003)
7 E. Marshall, Gene therapy's growing pains, Science, 269, 1050-1055 (1995)   DOI   ScienceOn
8 T. Niidome and L. Huang, Gene therapy progress and prospects: nonviral vectors, Gene Ther., 9, 1647-1652 (2002)   DOI   ScienceOn
9 C. Plank, K. Mechtler, F.C.J. Szoka and E. Wagner, Activation of the complement system by synthetic DNA complexes: a potential barrier for intravenous gene delivery, Hum. Gene Ther., 7, 1437-1446 (1996)   DOI   ScienceOn
10 P. Erbacher, J.S. Remy and IP. Behr, Gene transfer with synthetic virus-like particles via the integrin-mediated endocytosis pathway, Gene Ther., 6, 138-145 (1999)   DOI   ScienceOn
11 Y.K. Oh, D. Suh, J.M. Kim, H.G. Choi, K Shin and J.J. Ko, Polyethylenimine-mediated cellular uptake, nucleus trafficking and expression of cytokine plasmid DNA, Gene Ther., 9, 1627-1632 (2002)   DOI   ScienceOn
12 D. Lechardeur, K.J. Sohn, M. Haardt, P.E. Joshi, M. Monck, R.W. Graham, B. Beatty, J. Squire, H. Q'Brodovich and G.L. Lukacs, Metabolic instability of plasmid DNA in the cytosol: A potential barrier to gene transfer, Gene Ther., 6, 482-497 (1999)   DOI   ScienceOn
13 C.H. Ahn, S.Y. Chae, Y.H. Bae and S.W Kim, Biodegradable poly(ethylenimine) for plasmid DNA delivery, J. Controlled ReI., 80, 273-282 (2002)   DOI   ScienceOn
14 Y.K. Qh, IP. Kim, H. Yoon, J.M. Kim, J.S. Yang and C.K Kim, Prolonged organ retention and safety of plasmid DNA administered in polyethylenimine complexes, Gene Ther., 8, 1587-1592 (2001)   DOI   ScienceOn
15 A.R. Klemm, D. Young and J.B. Lloyd, Effects of polyethyl-eneimine on endocytosis and lysosome stability, Biochem. Plarmacol., 56, 41-46 (1998)   DOI   ScienceOn
16 F. Denizot and R Lang, Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability, J. Immunol. Methods, 89, 271-277 (1986)   DOI   ScienceOn
17 W.T. Godbey, K.K. Wu and A.G. Mikos, Size matters: Molecular weight affects the efficiency of poly(ethylenimine) as a gene delivery vehicle, J. Biomed. Mater. Res., 45, 268-275 (1999)   DOI   ScienceOn
18 W.T. Godbey, K.K. Wu and AG. Mikos, Poly(ethylenimine) and its role in gene delivery, J. Control Release, 60, 149-160 (1999)   DOI   ScienceOn