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

超高压制备的氢键互连聚合物组装体的基因递送应用

• 批准号: 16300167
• 负责人: KISHIDA Akio
• 金额: $ 7.36万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16300167/
• 关键词: hydrogen bond; high pressure; gene delivery; non-viral vector; inorganic particle; polymer; 遺伝子ベクター; ポリビニルアルコール; DNA

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.

综述了利用超高压组装分子技术制备新基因传递复合物的研究进展。今年，我们研究了以下主题：1)与聚乙烯醇以外的聚合物组装DNA的研究，2)具有氢键的无机微粒的应用。以葡聚糖、聚（丙烯酰胺）、聚（丙烯酸）、聚（乙二醇）和低分子量聚（甲基丙烯酸羟甲基酯）为氢键聚合物，考察了与DNA的组装。只有右旋糖酐与聚乙二醇的结合使用超高压工艺进行分子组装。研究了这两种聚合物与质粒DNA的组装。澄清的是，组装的形成在很大程度上受溶液状态的影响。空气中的二氧化碳在蒸馏水中溶解一次后，化合物就被阻挡了。在使用氢键比聚乙烯醇弱的聚合物的情况下，很明显，实验条件应该设定得更严格。检测了无机微粒表面与羟基结合的DNA。制备了羟基磷灰石纳米晶体，其表面具有羟基，在溶液中保持分布，并检测了纳米晶体与DNA的组装。DNA化合物稳定下来，并能够进行基因传递到细胞中的实验。质粒DNA的掺入量高，但编码蛋白的出现量低。这些结果表明，必须考虑一个新的机制，关于运输到细胞核的细胞和DNA与其他材料的组装条件的检查。由于低分子量肽的结构似乎不受超高压的影响，因此作为候选材料将被研究。