磷酸化壳聚糖-季铵化壳聚糖载体提高体内外基因转染功效

发布时间：2019-02-13 03:49

【摘要】：本研究通过磷酸化壳聚糖(PC)包被季铵化壳聚糖(TMC)/质粒(pDNA)纳米复合物,制备PC/TMC/pDNA三元复合物(PTC),用于提高体内外基因转染功效.PTC粒径为100~200nm,Zeta电势为-16~-34mV,可有效缩合pDNA,保护pDNA免遭核酶降解.PC降低PTC中pDNA结合力,增加体外释放量.表面荷负电的PTC抗非特异性蛋白吸附能力强,经小窝蛋白介导的细胞内吞途径入胞后逃避溶酶体降解,细胞核内分布比例高.HEK293细胞体外转染试验结果显示,PTC体外转染效率是TMC/pDNA纳米复合物(TC)的1.5~3.1倍;小鼠胫前肌注射给药试验结果表明,PTC可显著提高基因体内转染效率.因此,合适质量比的PTC有望作为功能性基因药物的递送载体,用于基因治疗.
[Abstract]:In this study, PC/TMC/pDNA ternary complex (PTC), was prepared by coating quaternary chitosan (TMC) / plasmid (pDNA) nanocomposites with phosphorylated chitosan (PC). The PC/TMC/pDNA ternary complex (PTC), was used to improve gene transfection efficiency in vivo and in vitro. Zeta potential was -16 ~ 34mV, which could effectively condense pDNA, to protect pDNA from ribozyme degradation. PC reduced pDNA binding force in PTC and increased release amount in vitro. PTC with negative surface charge has strong ability to resist non-specific protein adsorption, and escapes lysosome degradation through the intracellular endocytosis pathway mediated by nest protein. The distribution of lysosome in the nucleus is high. The results of HEK293 cell transfection in vitro showed that, The transfection efficiency of PTC was 1.5 ~ 3.1 times higher than that of TMC/pDNA nanocomposite (TC) in vitro. The results of tibialis anterior muscle injection test showed that PTC could significantly improve the efficiency of gene transfection in vivo. Therefore, PTC with appropriate mass ratio is expected to be used as a delivery vector for functional gene drugs for gene therapy.
【作者单位】：复旦大学生命科学学院遗传工程国家重点实验室;
【分类号】：R450

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