PLL/pDNA/P(His-co-DMAEL) ternary complexes: assembly, stability and gene delivery
作者:Dezhong Zhou、Congxin Li、Yuling Hu、Hao Zhou、Jiatong Chen、Zhengpu Zhang、Tianying Guo
DOI:10.1039/c2jm30850a
日期:——
Poly(L-lysine) (PLL) is a kind of biocompatible and biodegradable polycation and can effectively condense plasmid DNA (pDNA) to form nano-scaled complexes. However, the ultra-low gene transfection efficiency limits its wide application in gene delivery. Here, an uncharged functional polymer P(His-co-DMAEL) is synthesized by reversible addition fragmentation transfer (RAFT) polymerization and characterized with 1H NMR spectrum and size exclusion chromatography (SEC). P(His-co-DMAEL) is assembled with pDNA and PLL to form PLL/pDNA/P(His-co-DMAEL) ternary complexes. Gel shift assays and dynamic light scattering (DLS) tests indicate that the stability of PLL/pDNA/P(His-co-DMAEL) ternary complexes under various conditions is much better than that of PLL/pDNA binary complexes. In vitro transfection shows that the gene transfection efficiency of PLL/pDNA/P(His-co-DMAEL) ternary complexes is much higher than that of PLL/pDNA counterparts and the “gold standard” of gene transfection, poly(ethyleneimine)/pDNA (PEI/pDNA) complexes, especially under serum conditions. Cytotoxicity assays show that PLL/pDNA/P(His-co-DMAEL) ternary complexes have no cytotoxicity. The ternary complexes have advanced the application of PLL as a gene carrier and will have enlightening significance for the development of novel gene and drug delivery carriers.
聚(L-赖氨酸) (PLL) 是一种生物相容性、可生物降解的聚阳离子,可以有效凝聚质粒DNA (pDNA) 形成纳米级复合物。然而,超低的基因转染效率限制了其在基因递送中的广泛应用。在此,通过可逆加成断裂转移 (RAFT) 聚合合成了不带电荷的功能聚合物 P(His-co-DMAEL),并通过 1 H NMR 谱和尺寸排阻色谱 (SEC) 进行了表征。 P(His-co-DMAEL)与pDNA和PLL组装形成PLL/pDNA/P(His-co-DMAEL)三元复合物。凝胶位移实验和动态光散射(DLS)测试表明PLL/pDNA/P(His-co-DMAEL)三元复合物在各种条件下的稳定性均明显优于PLL/pDNA二元复合物。体外转染结果表明,PLL/pDNA/P(His-co-DMAEL)三元复合物的基因转染效率远高于PLL/pDNA对应物,是基因转染的“金标准”聚(乙烯亚胺)/pDNA (PEI/pDNA) 复合物,尤其是在血清条件下。细胞毒性实验表明PLL/pDNA/P(His-co-DMAEL)三元复合物没有细胞毒性。该三元配合物推进了PLL作为基因载体的应用,对新型基因和药物递送载体的开发具有启发意义。