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Int. J. Mol. Sci. 2007, 8(2), 81-102; doi:10.3390/i8020081

Efficient Gene Transfection into Mammalian Cells Mediated by Cross-linked Polyethylenimine

1 and 1,*
1 The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing 210093, Jiangsu, P. R. China 2 The School of Chemical Engineering, Nanjing University of Science and Technology, Nanjing 210094, Jiangsu, P. R. China 3 College of Basic Medical Science , Southeast University, Nanjing 210009, Jiangsu, P. R. China
* Author to whom correspondence should be addressed.
Received: 15 December 2006 / Accepted: 31 January 2007 / Published: 14 February 2007
(This article belongs to the Special Issue Interaction of Biological Molecules)
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25 kDa branched polyethylenimine (PEI) has successfully been used for in vitroand in vivo gene delivery approaches, but it is cytotoxic. Smaller PEIs are usually non-cytotoxic but less efficient. In order to enhance the gene delivery efficiency and minimizecytotoxicity of PEI, we explored to synthesize cross-linked PEIs with degradable bonds byreacting amines of small branched 2000 Da PEI with small diacrylate (1,4-butanedioldiacrylate or ethyleneglycol dimethacrylate) for 2-6 hours. The efficiency of the cross-linkedPEIs during in vitro delivering plasmid containing enhanced green fluorescent protein(EGFP) gene reporter and their cytotoxicity were assessed in melanoma B16F10 cell andother cell lines. In vivo gene delivery efficiency was evaluated by direct injection delivery ofthe EGFP plasmid/ cross-linked PEI complexes into mice and by estimating the EGFPexpression in animal muscles. Compared to commercially available 25-kDa branched PEI,the cross-linked PEIs reported here could mediate more efficient expression of reporter genethan the 25-kDa PEI control, 19-fold more efficiently in B16F10 cells, 17-fold in 293T cells, 2.3-fold in 3T3 cells, and they exhibited essentially nontoxic at their optimized condition for gene delivery. Furthermore the transfection activity of polyplexs was preserved in the presence of serum proteins. The muscle transfected with the cross-linked PEI prepared here exhibited normal morphology and excellent gene expression. The cross-linked PEIs reported here were evidently more efficient than the commercial 25-kD PEI control and had less cytotoxicity in gene delivery in vitro and in vivo.
Keywords: Polyethylenimine; Gene delivery; Degradable; Cytotoxicity; B16F10; Mouse muscle. Polyethylenimine; Gene delivery; Degradable; Cytotoxicity; B16F10; Mouse muscle.
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Dong, W.; Li, S.; Jin, G.; Sun, Q.; Ma, D.; Hua, Z. Efficient Gene Transfection into Mammalian Cells Mediated by Cross-linked Polyethylenimine. Int. J. Mol. Sci. 2007, 8, 81-102.

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