In vitro transfection of plasmid DNA by cationized gelatin prepared from different amine compounds.
The objective of this paper is to compare the in vitro transfection efficiency of a luciferase plasmid DNA using cationized gelatin prepared from different amine compounds. The compounds used here were ethylenediamine, putrescine, spermidine and spermine, chemically introduced to the carboxyl group of gelatin for the cationization. Complexation of the cationized gelatin with the plasmid DNA was performed by simply mixing the two materials at various N+/P- mixing ratios (the molar number ratio of amino groups of gelatin to the phosphate groups of DNA) in aqueous solution. Gel retardation studies revealed that the formation of cationized-gelatin-plasmid DNA complexes depended on the N+/P- mixing ratio. The stronger interaction of plasmid DNA with the cationized gelatin of spermine compared to the other cationized gelatins was observed by an ethidium bromide intercalation assay and Scatchard binding analysis. When the transfection efficiency of plasmid DNA complexed with the various cationized gelatins at different N+/P- mixing ratios was evaluated for mouse L929 fibroblasts, the highest transfection efficiency was observed for the complex prepared from the cationized gelatin of spermine at a N+/P- mixing ratio of 2. The present study indicates that there is an optimal N+/P- mixing ratio and a type of amine compound or cationization extent of cationized gelatin to enhance the transfection efficiency of plasmid DNA.
Kushibiki
Toshihiro
T
Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Tomoshige
Ryuji
R
Iwanaga
Kazunori
K
Kakemi
Masawo
M
Tabata
Yasuhiko
Y
D000588:Amines / Q000737:chemistry; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry* / Q000302:isolation & purification; D002412:Cations; D002460:Cell Line; D005780:Gelatin / Q000737:chemistry* / Q000302:isolation & purification; D015870:Gene Expression; D017930:Genes, Reporter; D049409:Luciferases, Firefly / Q000235:genetics; D008422:Materials Testing; D051379:Mice; D010957:Plasmids / Q000235:genetics*; D014162:Transfection
D000588:Amines; D001672:Biocompatible Materials; D002412:Cations; D005780:Gelatin; D049409:Luciferases, Firefly