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纤维连接蛋白-DNA-磷灰石复合层用于高效和特定区域的基因转染。

Fibronectin-DNA-apatite composite layer for highly efficient and area-specific gene transfer.

机构信息

Nanotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.

出版信息

J Biomed Mater Res A. 2010 Mar 1;92(3):1038-47. doi: 10.1002/jbm.a.32449.

Abstract

In 2004, Shen et al. developed a safe and efficient gene transfer system using a DNA-apatite composite layer. We have recently succeeded in improving further the gene transfer efficiency by immobilizing a cell adhesion molecule laminin, in a DNA-apatite composite layer. In this study, we showed that not only laminin but fibronectin immobilized in a DNA-apatite composite layer enhances cell adhesion and cell spreading on the layer, thereby markedly improving the gene transfer efficiency. Therefore, the immobilization of a cell adhesion molecule in a DNA-apatite composite layer is crucial for improving the gene transfer efficiency. By using fibronectin instead of laminin and optimizing the condition to prepare the fibronectin-DNA-apatite composite layer, the amount (weight) of cell adhesion molecule required was reduced to approximately one-fourth while retaining the relatively high gene transfer efficiency. It was also shown that the resulting fibronectin-DNA-apatite composite layer prepared under the optimized condition mediated the area-specific gene transfer on its surface, that is, DNA was preferentially transferred to the cells adhering to the surface of the fibronectin-DNA-apatite composite layer. The present gene transfer system with potential for area-specific transfection and advantages of safety and relatively high efficiency would be useful in tissue engineering applications, gene therapy, and production of transfection microarrays.

摘要

2004 年,Shen 等人开发了一种使用 DNA-磷灰石复合材料层的安全有效的基因转移系统。我们最近成功地通过在 DNA-磷灰石复合材料层中固定细胞黏附分子层粘连蛋白进一步提高了基因转移效率。在这项研究中,我们表明,不仅层粘连蛋白,而且固定在 DNA-磷灰石复合材料层中的纤连蛋白都能增强细胞在该层上的黏附和扩展,从而显著提高基因转移效率。因此,细胞黏附分子在 DNA-磷灰石复合材料层中的固定对于提高基因转移效率至关重要。通过使用纤连蛋白代替层粘连蛋白并优化制备纤连蛋白-DNA-磷灰石复合材料层的条件,所需细胞黏附分子的量(重量)减少到大约四分之一,同时保持相对较高的基因转移效率。还表明,在优化条件下制备的纤连蛋白-DNA-磷灰石复合材料层介导了其表面的区域特异性基因转移,即 DNA 优先转移到黏附在纤连蛋白-DNA-磷灰石复合材料层表面的细胞上。具有区域特异性转染潜力且具有安全和相对高效优点的本基因转移系统将在组织工程应用、基因治疗和转染微阵列的制备中非常有用。

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