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Research Article

短合成脂肽/Importin-α的计算机分子相互作用和基于脂质体的基因载体介导的转基因表达的体外评估

卷 20, 期 5, 2020

页: [383 - 394] 页: 12

弟呕挨: 10.2174/1566523220666201005104224

价格: $65

摘要

背景：基于脂肽的基因载体显示出低细胞毒性，能够穿透细胞膜，易于制造，因此是基因递送应用的巨大潜力。 目的：本研究旨在探索一系列由烷基链，一个半胱氨酸（C），1-2个组氨酸（H）和赖氨酸（K）残基组成的短合成脂肽（Lau：Lauryl； Pal：Palmitoyl）。进行计算机内分子相互作用和体外评估。 方法：使用AutoDock Vina和Amber14进行脂肽和Importin-α受体之间的分子相互作用。体外评估脂肽/ DNA复合物的相互作用，粒径，ζ电势和转基因表达。基于萤光素酶转基因表达，进行脂肽和Pal-CKKHH来源的脂质体的转染效率。 结果：计算机内相互作用表明，Lau-CKKH和Pal-CKKHH假设加速了核吸收。与天然配体，即核定位序列（-5.4kcal / mol）相比，两种脂肽均具有较低的结合能（分别为-6.3kcal / mol和-6.2kcal / mol）。短的脂肽能够浓缩DNA分子并有效形成紧密的纳米颗粒。根据对COS-7的体外评估，发现Pal-CKKHH是脂肽中最好的转染剂。与辅助脂质DOPE（1：2）一起配制时，其转染效率（ng Luc / mg总蛋白）提高了约3倍（1163 + 55）。基于脂肽的脂质体（Pal-CKKHH：DOPE = 1：2）还促进了萤光素酶转基因在人胚胎肾细胞（293T）和人宫颈腺癌细胞（HeLa）上的表达，转染效率分别为1779 +52和260 + 22。 结论：我们的首次研究表明，完全合成的短脂肽Pal-CKKHH能够与Importin-α牢固地相互作用。脂肽能够有效地凝结DNA分子，促进转基因表达，加速核摄取过程，因此具有潜在的转染剂的特征。

关键词: 脂肽，importin-α，计算机内，脂质体，粒径，转染。

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图形摘要

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