Gene delivery application of the polymer assembly interconnected by hydrogen bonding prepared by using ultra-high pressure

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The development of the complex for new gene delivery by the assembling molecule technology that utilizes the ultra high pressure was examined. This year, we examined following themes: 1) Examination of assembling DNA with polymers other than polyvinyl alcohol, 2) Application of inorganic micro particle that has hydrogen bond. The assembly with DNA was examined by using the polysaccharide including dextran, poly (acrylamide), poly (acrylic acid), poly (ethyleneglycol), and low molecular weight poly (hydroxymethylmethacrylate) as hydrogen bond polymers. Only the combination of dextran with poly (ethyleneglycol) gave molecular assembly using ultra-high pressure process. The assembly with plasmid DNA was examined about these two kinds of polymers. It was clarified that the formation of assembly was greatly influenced by the state of the solution. The compound was obstructed about the one that carbon dioxide in air dissolved to the distilled water once. In case of using the polymer to which the hydrogen bonds are weaker than poly (vinyl alcohol), it became clear that the experimental condition should be set more strictly. The assembling DNA with the hydroxyl group on the surface of the inorganic micro particle was examined. The hydroxyapatite nanocrystal that kept distributing in solution and had the hydroxyl groups on its surface was prepared, and the assembling with nanocrystals and DNA was examined. The compound of DNA stabilized, and was able to conduct the experiment on the gene delivery into the cell. The incorporation of plasmid DNA was high, however, the amount of appearance of the code protein was low. These results showed that a new mechanism had to be considered about the transportation to the nucleus in the cell and the examination of the condition of DNA assembling with other materials. As the candidate material, low molecular weight peptide will be examined because of that the structure of the peptide seemed not to be affected by the ultra-high pressure.