Chowdhury E H, Akaike Toshihiro
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Biotechnol Bioeng. 2005 May 20;90(4):414-21. doi: 10.1002/bit.20398.
The DNA delivery to mammalian cells is an essential tool for analyzing gene structure, regulation, and function. The approach holds great promise for the further development of gene therapy techniques and DNA vaccination strategies to treat and control diseases. Here, we report on the establishment of a cell-specific gene delivery and expression system by physical adsorption of a cell-recognition molecule on the nano-crystal surface of carbonate apatite. As a model, DNA/nano-particles were successfully coated with asialofetuin to facilitate uptake by hepatocyte-derived cell lines through the asialoglycoprotein receptor (ASGPr) and albumin to prevent non-specific interactions of the particles with cell-surface. The resulting composite particles with dual surface properties could accelerate DNA uptake and enhance expression to a notable extent. Nano-particles coated with transferrin in the same manner dramatically enhanced transgene expression in the corresponding receptor-bearing cells and thus our newly developed strategy represents a universal phenomenon for anchoring a bio-recognition macromolecule on the apatite crystal surface for targeted gene delivery, having immediate applications in basic research laboratories and great promise for gene therapy.
将DNA导入哺乳动物细胞是分析基因结构、调控和功能的重要工具。这种方法对于基因治疗技术和DNA疫苗接种策略的进一步发展具有巨大潜力,有望用于治疗和控制疾病。在此,我们报告了一种通过在碳酸磷灰石纳米晶体表面物理吸附细胞识别分子来建立细胞特异性基因递送和表达系统的方法。作为模型,DNA/纳米颗粒成功地用去唾液酸胎球蛋白包被,以促进肝细胞衍生细胞系通过去唾液酸糖蛋白受体(ASGPr)摄取,并使用白蛋白防止颗粒与细胞表面发生非特异性相互作用。所得具有双重表面性质的复合颗粒可显著加速DNA摄取并增强表达。以同样方式用转铁蛋白包被的纳米颗粒显著增强了相应受体细胞中的转基因表达,因此我们新开发的策略代表了一种将生物识别大分子锚定在磷灰石晶体表面以进行靶向基因递送的普遍现象,在基础研究实验室中有直接应用,对基因治疗也有巨大潜力。