Efficient gene delivery of primary human cells using peptide linked polyethylenimine polymer hybrid.
Efficient gene delivery of primary human cells using peptide linked polyethylenimine polymer hybrid.
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利用肽连接聚乙烯亚胺聚合物杂交体向原代人体细胞高效传递基因。
DOI：
10.1016/j.biomaterials.2011.03.016
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发表时间：
2011-07
期刊：
影响因子：
14
通讯作者：
Wu, Joseph C.
中科院分区：
文献类型：
--
作者：
Dey, Devaveena;Inayathullah, Mohammed;Lee, Andrew S.;LeMieux, Melburne C.;Zhang, Xuexiang;Wu, Yi;Nag, Divya;De Almeida, Patricia Eliza;Han, Leng;Rajadas, Jayakumar;Wu, Joseph C.
基于聚乙烯亚胺 (PEI) 的聚合物是细胞转染的有效试剂。然而，由于与转染相关的高细胞死亡，从而导致细胞内基因递送效率低，因此它们的使用受到阻碍。为了规避细胞毒性问题，将金属结合肽与 PEI 连接。合成了八种肽-PEI 衍生物以提高细胞存活率和转染效率。 TAT 连接的 PEI 用作对照聚合物。电子显微镜和原子力显微镜测量显示，与 PEI 胺连接的肽形成纳米凝胶。通过光谱方法对聚合物进行表征，并使用电泳研究测试它们与质粒形成复合物的能力。与单独的 PEI 相比，这些修饰显着改善了聚合物的生物相容性以及细胞存活率。与广泛使用的转染剂 lipofectamine 相比，修饰的肽聚合物的一个子集在原代人类细胞中也显示出显着更高的转染效率。对所观察到的现象的潜在机制的研究表明，与单独使用 PEI 相比，肽聚合物存在下的“活性氧”(ROS) 水平较低。全局基因表达分析进一步证实了这一点，该分析显示参与调节细胞内氧化应激的多个基因和途径上调。
Polyethylenimine (PEI) based polymers are efficient agents for cell transfection. However, their use has been hampered due to high cell death associated with transfection thereby resulting in low efficiency of gene delivery within the cells. To circumvent the problem of cellular toxicity, metal binding peptides were linked to PEI. Eight peptide-PEI derivatives were synthesized to improve cell survival and transfection efficiency. TAT linked PEI was used as a control polymer. Peptides linked with PEI amines formed nano gels as shown by electron microscopy and atomic force microscopic measurements. Polymers were characterized by spectroscopic methods and their ability to form complexes with plasmids was tested using electrophoretic studies. These modifications improved polymer biocompatibility as well as cell survival markedly when compared to PEI alone. A subset of the modified peptide-polymers also showed significantly higher transfection efficiency in primary human cells with respect to the widely used transfection agent, lipofectamine. Study of the underlying mechanism of the observed phenomena revealed lower levels of ‘reactive oxygen species’ (ROS) in presence of the peptide-polymers when compared to PEI alone. This was further corroborated with global gene expression analysis which showed upregulation of multiple genes and pathways involved in regulating intracellular oxidative stress.
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