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.
Highlights
The development of gene delivery systems is an important area in the field of genetic engineering [1]
The fact that the efficiency increased in a concentration-dependent manner may be due to an increase in transfection efficiency resulting from low DDMC cell toxicity for their increases in the complex formed with DNA
DDMC, which is used as a carrier for gene introduction, can be sterilized by autoclaving, has better transfection efficiency relative to DEAE-dextran alone, and is thought to have lower cellular toxicity
Summary
The development of gene delivery systems is an important area in the field of genetic engineering [1]. A constituent element involves the transport of genes, which requires a transport vehicle referred to as a vector. Vectors include viral “shells” or lipid spheres (liposomes) having properties whereby they are incorporated into host cells. Liposome vectors are completely artificial and are produced by introducing genes into microspheres that have a lipid bilayer structure similar to that of a cell membrane. Favorable results regarding efficiency have been indicated with commercial cationic lipid micelle transfection reagents. These reagents cannot be sterilized by autoclaving and are not amenable to mainstream use as non-viral gene introduction carriers. Electrophoresis and microinjection methods are examples of electrical and physical methods, but they require special devices and technologies
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