110(P-54) カチオン性フラーレン集合体による新規遺伝子導入法の開発(ポスター発表の部)

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We previously reported that an aminofullerene can deliver extra-cellular gene into mammalian cells, and that an appropriately designed amphiphilic fullerene functions as a new transfection reagent. In the present study, we found that the aminofullerene acts as a potent transfection reagent for the long-term expression of the gene. The aminofullerene protects the extracellular DNA from the nuclease digestion, which provides the key for the successful stable transfection. We also report synthesis of a newly designed fullerene transfection reagent. Using a new method for the synthesis of tetraaminofullerene epoxides, the "second generation" reagent was prepared in good yield by two-step transformation of C_<60>. We used an ordinary stable transfection assay for the evaluation of the long-term expression. Thus, a plasmid DNA carrying hygromysin B resistance gene was transfected and the efficiency was determined after two-week selective growth of COS-1 cells. The long-term transfection efficiency of fullerene 1 was 0.73%, which is 7 times higher than that of a control experiment using lipofectin (0.10%). By using cellular assay and test-tube assay, we found that aminofullerene protect DNA from the digestion by nuclease. In the digestion experiment of plasmid DNA with endonuclease, we found that the hydrolysis by endonuclease was completely inhibited in the presence of fullerene 1, whereas lipofectin protects DNA only partially. Development of "second generation" fullerene transfection reagent will also be reported.
天然有機化合物討論会の論文
2004-10-01

110(P-54) カチオン性フラーレン集合体による新規遺伝子導入法の開発(ポスター発表の部)

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