Novel guanidinylated bioresponsive poly(amidoamine)s designed for short hairpin RNA delivery.
Novel guanidinylated bioresponsive poly(amidoamine)s designed for short hairpin RNA delivery.
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新型胍基化生物响应性聚酰胺胺，专为短发夹 RNA 递送而设计
DOI：
10.2147/ijn.s115773
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发表时间：
2016
影响因子：
8
通讯作者：
Ding P
中科院分区：
文献类型：
--
作者：
Yu J;Zhang J;Xing H;Sun Y;Yang Z;Yang T;Cai C;Zhao X;Yang L;Ding P
以含胍(GUA)的单体(精氨酸[Arg]和胍丁胺[AGM])与N，N‘-半胱胺双丙烯酰胺(CBA)为原料，通过Michael-加成聚合反应合成了两种不同类型的含二硫键(SS)的聚酰胺(PAA)聚合物。为了表征这两种GUA-SS-PAA聚合物及其作为短发夹状RNA(ShRNA)载体的潜力，选择pSilencer 4.1-CMV FANCFshRNA作为DNA模型与这两种聚合物形成复合体。在核酸：聚合物质量比小于1：24的条件下，测得GUA-SS-PAAS和质粒DNA的ζ-电位均小于20 mV，粒径小于90 nm。从两个新构建的复合体中都观察到了有生物反应性的DNA释放。与作为参比载体的两种广泛使用的转染剂聚乙烯亚胺和脂质体2000相比，两种聚合物复合体的细胞毒性都显著降低。Arg-CBA在MCF7细胞中的转染率和基因沉默效率均高于AGM-CBA和参比载体。此外，MCF7细胞对Arg-CBA的摄取高于AGM-CBA和参比载体。同样，Arg-CBA介导的DNA进入细胞核的能力强于AGM-CBA和参比载体。研究表明，在GUA-SS-PAAS聚合物中引入胍基和羧基具有更好的核定位效果，这在观察到的提高转染率和降低细胞毒性方面起到了关键作用。总体而言，两种新合成的GUA-SS-PAAS聚合物显示出作为shRNA载体用于基因治疗应用的巨大潜力。
Two different disulfide (SS)-containing poly(amidoamine) (PAA) polymers were constructed using guanidino (Gua)-containing monomers (ie, arginine [Arg] and agmatine [Agm]) and N,N′-cystamine bisacrylamide (CBA) by Michael-addition polymerization. In order to characterize these two Gua-SS-PAA polymers and investigate their potentials as short hairpin RNA (shRNA)-delivery carriers, pSilencer 4.1-CMV FANCF shRNA was chosen as a model plasmid DNA to form complexes with these two polymers. The Gua-SS-PAAs and plasmid DNA complexes were determined with particle sizes less than 90 nm and positive ζ-potentials under 20 mV at nucleic acid:polymer weight ratios lower than 1:24. Bioresponsive release of plasmid DNA was observed from both newly constructed complexes. Significantly lower cytotoxicity was observed for both polymer complexes compared with polyethylenimine and Lipofectamine 2000, two widely used transfection reagents as reference carriers. Arg-CBA showed higher transfection efficiency and gene-silencing efficiency in MCF7 cells than Agm-CBA and the reference carriers. In addition, the cellular uptake of Arg-CBA in MCF7 cells was found to be higher and faster than Agm-CBA and the reference carriers. Similarly, plasmid DNA transport into the nucleus mediated by Arg-CBA was more than that by Agm-CBA and the reference carriers. The study suggested that guanidine and carboxyl introduced into Gua-SS-PAAs polymers resulted in a better nuclear localization effect, which played a key role in the observed enhancement of transfection efficiency and low cytotoxicity. Overall, two newly synthesized Gua-SS-PAAs polymers demonstrated great potential to be used as shRNA carriers for gene-therapy applications.