Hepatocyte gene transfer mediated by stable polyplexes based on MPP-containing DNA complexes

• Published in: Hepatobiliary & pancreatic diseases international Vol. 8; no. 5; pp. 498 - 503
• Main Authors: Yu, Bao-Feng, Li, Wan-I, Hu, Xiao-Nian, Zhang, Yue-Hong, Niu, Bo, Xie, Jun
• Format: Journal Article
• Language: English
• Published: Singapore 01.10.2009
• Subjects: Animals; Cells, Cultured; DNA - metabolism; gene; Gene Transfer Techniques; Genetic Therapy - methods; Genetic Vectors; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; hepatocyte; Hepatocytes - cytology; Hepatocytes - metabolism; Histidine - genetics; Histidine - metabolism; Liver - cytology; Liver - metabolism; Male; membrane; Mice; Mice, Inbred BALB C; Models, Animal; penetrating; peptide; Plasmids; protein; therapy; transduction
• ISSN: 1499-3872

BACKGROUND:In the field of gene therapy,viral vectors as delivery tools have a number of disadvantages for medical application.This study aimed to explore a novel nonviral vector as a vehicle for gene therapy. METHODS:Transvector-rpE-MPP and EGFP(enhanced green fluorescent protein)were used as the gene transfer carrier and the reporter gene,respectively.Polyplexes which integrate transvector-rpE-MPP,the object gene,and EGFP were formed.The optimal charge ratio,stability, and transduction capacity of the polyplexes in mouse hepatocytes in vitro and in mouse liver in vivo were investigated.The polyplexes of transvector-rpE-MPP and pcDNA3-EGFP,with charge ratios of 0,0.25,0.5,0.75,1 and 1.5 were compared to determine the optimal charge ratio. RESULTS:Polyplexes with charge ratios of 1∶1 were most stable;pcDNA3-EGFP in these complexes resisted digestion by DNaseⅠand blood plasma.On the other hand,pcDNA3-EGFP alone was digested.Fluorescence analysis indicated that transvector-rpE-MPP successfully delivered the reporter gene EGFP into hepatocytes and that EGFP expression was detected in hepatocyte cultures and in liver tissue. CONCLUSION:These results have laid a foundation for further study of a novel nonviral gene delivery system.