鱼精蛋白对寡核苷酸复合物的缩合作用及其对转染效果的影响

Membrane Permeable Peptide-Protamine-Triplex forming Oligodeoxinucleotide Nanoparticles Transfect the Human Umbilical Vein Endothelial cell

目的观察鱼精蛋白对寡核甘酸复合物的缩合效果。方法用不同浓度鱼精蛋白对寡核甘酸进行缩合,凝胶电泳和扫描电镜观测其缩合结果。将设计的双功能肽与鱼精蛋白和寡核甘酸缩合物进行不同浓度配比后转染脐静脉内皮细胞,激光共聚焦显微镜下观察寡核甘酸入胞效果。结果在鱼精蛋白与寡核甘酸质量比达到1.6∶1以上时能够明显促使寡核甘酸缩合,扫描电镜下观察发现优于脂质体LipofetamineTM2000对该寡核甘酸的缩合。当功能肽与鱼精蛋白和寡核甘酸缩合物以质量比1∶5-1∶10配比复合时能够促使寡核甘酸进入脐静脉内皮细胞且转染率较高。结论优化的双功能肽-鱼精蛋白-寡核苷酸复合法,能够有效地缩合寡核苷酸,并促进寡核甘酸向脐静脉内皮细胞胞内转移。

Objective Optimize the nanoparticles construct with MPP-Protamine-oligodeoxinucleotide,to improve transfection efficiency that Triplex forming oligodeoxinucleotide into human umbilical vein endothelial cell.Methods Synthesize oligodeoxinucleotide labeled by FITC on the 5’ ends.Condensation the oligodeoxinucleotide with different matching Protamine that was evaluated by Gel retardation assay and transmission electron microscop.Construct with MPP-Protamine-oligodeoxinucleotide and observe their transfection efficiencies in HUVEC by laser scanning confocal microscopy.Results oligodeoxinucleotide can be Condensated into nanometer nanoparticles when the matching of Protamine get to 1.6.MPP match with Protamine-oligodeoxinucleotide 1∶10（weight）is higest transfection efficiencies.Conclusion MPP-Protamine-oligodeoxinucleotide is promising to provide a new safe carrier for the intracellular treatmen.