巯基烷基化壳聚糖载绿色荧光蛋白质粒基因纳米粒子的制备与研究

Preparation and Studies of Thiolated N-Alkylated Chitosan-Green Fluorescent Protein pDNA Nanoparticles

合成了新型的基因载体巯基烷基化壳聚糖(TACS),采用复凝聚法制备了巯基烷基化壳聚糖基因纳米粒子,并表征其形态和粒径。利用体外转染实验定性评估了纳米粒子的转染效率。结果表明,粒子形态不很均一,粒径在390nm左右;凝胶电泳实验证明了载体能有效地包裹和保护基因不受DNaseⅠ酶的消化;溶解实验证实了巯基化增强了TACS的水溶性;体外基因转染实验证实了TACS能够在人胚胎肾细胞(HEK293)转染基因,并且其转染效率远高于壳聚糖纳米粒子。这些结果表明,巯基烷基化壳聚糖有望成为有价值的基因载体。

Thiolated N-alkylated chitosan （TACS） was developed as a novel tool for gene delivery and the modified chitosan-pDNA nanoparticles were prepared by complex coacervation. The morphology of nanoparticles was irregular, observed by transmission electronic microscopy （TEM）. The average particle size was 390nm determined by nanoparticle size analyzer. The TACS-pDNA nanoparticles could partially protect the encapsulated plasmid pDNA from nuclease degradation as shown by electrophoretie mobility analysis. The solubility test showed sulfhedrylated reaction enhanced the water solubility of TACS. The gene transfeetion experiment in vitro suggested that the TACS-pDNA nanoparticles could transfect HEK293 effectively and the transfection efficiency of TACS-pDNA nanoparticles was much higher than that of chitosan-pDNA nanoparticles. All these identified TACS as a promising vector for gene delivery.