Mechanism of introduction of exogenous genes into cultured cells using DEAE-dextran-MMA graft copolymer as non-viral gene carrier.
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.
Eshita
Yuki
Y
Department of Infectious Disease Control, Faculty of Medicine, Oita University, 1-1 Idaigaoka, Hasama-machi, Yufu-shi, Oita Prefecture 879-5593, Japan. yeshita@med.oita-u.ac.jp
Higashihara
Junko
J
Onishi
Masayasu
M
Mizuno
Masaaki
M
Yoshida
Jun
J
Takasaki
Tomohiko
T
Kubota
Naoji
N
Onishi
Yasuhiko
Y
D000818:Animals; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D003911:Dextrans / Q000737:chemistry*; D008768:Methylmethacrylates / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
D003911:Dextrans; D008768:Methylmethacrylates; C544087:diethylaminoethyl-dextran-methyl methacrylate graft copolymer; D004247:DNA