Abstract: Comparative investigations were carried out regarding the efficiency of introduction of exogenous genes into cultured cells using a cationic polysaccharide DEAE-dextran-MMA (methyl methacrylate ester) graft copolymer (2-diethylaminoethyl-dextran-methyl methacrylate graft copolymer; DDMC) as a nonviral carrier for gene introduction. The results confirmed that the gene introduction efficiency was improved with DDMC relative to DEAE-dextran. Comparative investigations were carried out using various concentrations of DDMC and DNA in the introduction of DNA encoding luciferase (pGL3 control vector; Promega) into COS-7 cells derived from African green monkey kidney cells. The complex formation reaction is thought to be directly proportional to the transformation rate, but the complex formation reaction between DDMC and DNA is significantly influenced by hydrophobic bonding strength along with hydrogen bonding strength and Coulomb forces due to the hydrophobicity of the grafted MMA sections. It is thought that the reaction is a Michaelis-Menten type complex formation reaction described by the following equation: Complex amount = K1 (DNA concentration)(DDMC concentration). In support of this equation, it was confirmed that the amount of formed complex was proportional to the RLU value.
Keywords: transfection efficiency; DEAE-dextran-MMA graft copolymer; non-viral gene carrier; exogenous genes
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Eshita, Y.; Higashihara, J.; Onishi, M.; Mizuno, M.; Yoshida, J.; Takasaki, T.; Kubota, N.; Onishi, Y. Mechanism of Introduction of Exogenous Genes into Cultured Cells Using DEAE-Dextran-MMA Graft Copolymer as Non-Viral Gene Carrier. Molecules 2009, 14, 2669-2683.
Eshita Y, Higashihara J, Onishi M, Mizuno M, Yoshida J, Takasaki T, Kubota N, Onishi Y. Mechanism of Introduction of Exogenous Genes into Cultured Cells Using DEAE-Dextran-MMA Graft Copolymer as Non-Viral Gene Carrier. Molecules. 2009; 14(7):2669-2683.
Eshita, Yuki; Higashihara, Junko; Onishi, Masayasu; Mizuno, Masaaki; Yoshida, Jun; Takasaki, Tomohiko; Kubota, Naoji; Onishi, Yasuhiko. 2009. "Mechanism of Introduction of Exogenous Genes into Cultured Cells Using DEAE-Dextran-MMA Graft Copolymer as Non-Viral Gene Carrier." Molecules 14, no. 7: 2669-2683.