Ternary complexes of pDNA, polyethylenimine, and γ-polyglutamic acid for gene delivery systems
Ternary complexes of pDNA, polyethylenimine, and γ-polyglutamic acid for gene delivery systems
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DOI：
10.1016/j.biomaterials.2009.01.055
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发表时间：
2009-05-01
期刊：
影响因子：
14
通讯作者：
Sasaki, Hitoshi
中科院分区：
文献类型：
--
作者：
Kurosaki, Tomoaki;Kitahara, Takashi;Sasaki, Hitoshi
我们发现了一种涂有 γ-聚谷氨酸 (γ-PGA) 的载体，可实现有效且安全的基因传递。为了开发有用的非病毒载体，我们制备了几种由pDNA、聚乙烯亚胺（PEI）和各种聚阴离子构建的三元复合物，例如聚腺苷酸、聚肌苷-聚胞苷酸、α-聚天冬氨酸、α-聚谷氨酸和γ-PGA。 pDNA/PEI复合物具有强阳离子表面电荷，尽管与红细胞发生凝集并具有极高的细胞毒性，但仍表现出极高的转基因效率。这些聚阴离子将 pDNA/PEI 复合物的正 zeta 电位变为负值，但不影响大小。它们没有凝集活性并且细胞毒性较低，但大多数三元复合物没有表现出任何摄取和基因表达：然而，pDNA/PEI/gamma-PGA复合物表现出高摄取和基因表达。大多数 pDNA/PEI/gamma-PGA 复合物位于细胞质中，没有解离，少数复合物在细胞核中观察到。低温和添加γ-PGA显着抑制细胞对pDNA/PEI/γ-PGA的摄取，但L-谷氨酸没有影响。这些结果强烈表明pDNA/PEI/γ-PGA复合物被γ-PGA特异性受体介导的能量依赖性过程吸收。因此，pDNA/PEI/γ-PGA复合物可用作具有高转染效率和低毒性的基因递送系统。 (C) 2009 Elsevier Ltd. 保留所有权利。
We discovered a vector coated by gamma-polyglutamic acid (gamma-PGA) for effective and safe gene delivery. In order to develop a useful non-viral vector, we prepared several ternary complexes constructed with pDNA, polyethylenimine (PEI), and various polyanions, such as polyadenylic acid, polyinosinic-polycytidylic acid, alpha-polyaspartic acid, alpha-polyglutamic acid, and gamma-PGA. The pDNA/PEI complex had a strong cationic Surface charge and showed extremely high transgene efficiency although it agglutinated with erythrocytes and had extremely high cytotoxicity. Those polyanions changed the positive zeta-potential of pDNA/PEI complex to negative although they did not affect the size. They bad no agglutination activities and lower cytotoxicities bur most of the ternary complexes did not show any uptake and gene expression: however, the pDNA/PEI/gamma-PGA complex showed high uptake and gene expression. Most of the pDNA/PEI/gamma-PGA complexes were located in the Cytoplasm Without dissociation and a few complexes were observed in the nuclei. Hypothermia and the addition of gamma-PGA significantly inhibited the uptake of pDNA/PEI/gamma-PGA by the cells, although L-glutamic acid had no effect. These results strongly indicate that the pDNA/PEI/gamma-PGA complex was taken up by gamma-PGA-specific receptor-mediated energy-dependent process. Thus, the pDNA/PEI/gamma-PGA complex is useful as a gene delivery system with high transfection efficiency and low toxicity. (C) 2009 Elsevier Ltd. All rights reserved.