【摘要】：本研究通過磷酸化殼聚糖(PC)包被季銨化殼聚糖(TMC)/質(zhì)粒(pDNA)納米復(fù)合物,制備PC/TMC/pDNA三元復(fù)合物(PTC),用于提高體內(nèi)外基因轉(zhuǎn)染功效.PTC粒徑為100~200nm,Zeta電勢為-16~-34mV,可有效縮合pDNA,保護(hù)pDNA免遭核酶降解.PC降低PTC中pDNA結(jié)合力,增加體外釋放量.表面荷負(fù)電的PTC抗非特異性蛋白吸附能力強(qiáng),經(jīng)小窩蛋白介導(dǎo)的細(xì)胞內(nèi)吞途徑入胞后逃避溶酶體降解,細(xì)胞核內(nèi)分布比例高.HEK293細(xì)胞體外轉(zhuǎn)染試驗(yàn)結(jié)果顯示,PTC體外轉(zhuǎn)染效率是TMC/pDNA納米復(fù)合物(TC)的1.5~3.1倍;小鼠脛前肌注射給藥試驗(yàn)結(jié)果表明,PTC可顯著提高基因體內(nèi)轉(zhuǎn)染效率.因此,合適質(zhì)量比的PTC有望作為功能性基因藥物的遞送載體,用于基因治療.
[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.
【作者單位】： 復(fù)旦大學(xué)生命科學(xué)學(xué)院遺傳工程國家重點(diǎn)實(shí)驗(yàn)室;
【分類號(hào)】：R450

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