Application of an HIV gp41-derived peptide for enhanced intracellular trafficking of synthetic gene and siRNA delivery vehicles
Application of an HIV gp41-derived peptide for enhanced intracellular trafficking of synthetic gene and siRNA delivery vehicles
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DOI：
10.1021/bc700448h
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发表时间：
2008-04-01
影响因子：
4.7
通讯作者：
Pun, Suzie H.
中科院分区：
文献类型：
--
作者：
Kwon, Ester J.;Bergen, Jamie M.;Pun, Suzie H.
内体释放是许多非病毒基因递送载体的效率限制步骤。在这项工作中，非病毒基因递送载体用取自 HIV gp41 胞内结构域的膜溶解肽进行了修饰。肽与聚乙烯亚胺 (PEI) 共价连接，肽修饰的聚合物与 DNA 复合。所得纳米颗粒具有与未改性 PEI 形成的复合物相似的物理化学性质。 gp41 衍生肽无论是作为游离肽还是与 PEI 缀合时都表现出显着的裂解活性。与低聚合物与 DNA 比率的未修饰复合物相比，HeLa 细胞中转基因表达显着增加。此外，与未修饰的聚合复合物相比，肽修饰的聚合复合物介导的 siRNA 递送显着增强。尽管转基因表达和 siRNA 敲低有所增加，但根据流式细胞术测定，修饰载体的内化或结合没有增加。 gp41 衍生肽增加载体的内体逃逸的假设得到转染细胞中 DNA 分布的共焦显微镜成像的支持。这项工作展示了使用裂解肽来改善非病毒基因递送载体的运输。
Endosomal release is an efficiency-limiting step for many nonviral gene delivery vehicles. In this work, nonviral gene delivery vehicles were modified with a membrane-lytic peptide taken from the endodomain of HIV gp41. Peptide was covalently linked to polyethylenimine (PEI) and the peptide-modified polymer was complexed with DNA. The resulting nanoparticles were shown to have similar physicochemical properties as complexes formed with unmodified PEI. The gp41-derived peptide demonstrated significant lytic activity both as free peptide and when conjugated to PEI. Significant increases in transgene expression were achieved in HeLa cells when compared to unmodified polyplexes at low polymer to DNA ratios. Additionally, peptide-modified polyplexes mediated significantly enhanced siRNA delivery compared to unmodified polyplexes. Despite increases in transgene expression and siRNA knockdown, there was no increase in internalization or binding of modified carriers as determined by flow cytometry. The hypothesis that the gp41-derived peptide increases the endosomal escape of vehicles is supported by confocal microscopy imaging of DNA distributions in transfected cells. This work demonstrates the use of a lytic peptide for improved trafficking, of nonviral gene delivery vehicles.