Cross-linked polyethylenimine as potential DNA vector for gene delivery with high efficiency and low cytotoxicity.
Polyethylenimine (PEI) has been known as an efficient gene carrier with the highest cationic charge potential. High transfection efficiency of PEI, along with its cytotoxicity, strongly depends on its molecular weight. To enhance its gene delivery efficiency and minimize cytotoxicity, we have synthesized small cross-linked PEI with biodegradable linkages and evaluated their transfection efficiencies in vitro. In this study, branched PEI with a molecular weight of 800 Da was cross-linked by small diacrylate [1,4-butanediol diacrylate or ethyleneglycol dimethacrylate (EGDMA)] for 2-6 h. The efficiencies of the cross-linked PEI in in vitro transfection of plasmid DNA containing enhanced green fluorescent protein (EGFP) reporter gene were assessed in melanoma B16F10 cell line and other cell lines. Flow cytometry was used to quantify the cellular entry efficiency of plasmid and the transgene expression level. The cytotoxicities of the cross-linked PEI in these cells were evaluated by MTT assay. EGDMA-PEI 800-4h, a typical cross-linked PEI reported here, mediated a more efficient expression of reporter gene than the commercially available 25-kDa branched PEI control, and resulted in a 9-fold increase in gene delivery in B16F10 cells and a 16-fold increase in 293T cells, while no cytotoxicity was found at the optimized condition for gene delivery. Furthermore, the transfection activity of polyplexes was preserved in the presence of serum proteins.
Dong
Wei
W
State Key Laboratory of Pharmaceutical Biotechnology, College of Life Science, Nanjing University, Nanjing 210093, China.
Jin
Guang-Hui
GH
Li
Shu-Feng
SF
Sun
Qi-Ming
QM
Ma
Ding-Yuan
DY
Hua
Zi-Chun
ZC
D000818:Animals; D002072:Butylene Glycols / Q000737:chemistry*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004247:DNA / Q000008:administration & dosage*; D004337:Drug Carriers; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008689:Methacrylates / Q000737:chemistry*; D051379:Mice; D010957:Plasmids; D011094:Polyethyleneimine / Q000737:chemistry* / Q000633:toxicity
D002072:Butylene Glycols; D003432:Cross-Linking Reagents; D004337:Drug Carriers; D008689:Methacrylates; C485184:enhanced green fluorescent protein; C022288:butanediol diacrylate; D049452:Green Fluorescent Proteins; C004919:ethylene dimethacrylate; D011094:Polyethyleneimine; D004247:DNA