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首页> 外文期刊>Advanced Functional Materials >Chondroitin Sulfate-Coated DNA-Nanoplexes Enhance Transfection Efficiency by Controlling Plasmid Release from Endosomes: A New Insight into Modulating Nonviral Gene Transfection
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Chondroitin Sulfate-Coated DNA-Nanoplexes Enhance Transfection Efficiency by Controlling Plasmid Release from Endosomes: A New Insight into Modulating Nonviral Gene Transfection

机译：硫酸软骨素包裹的DNA纳米复合物通过控制质粒从内体释放来增强转染效率：调节非病毒基因转染的新见解。

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Degradation of plasmid DNA (pDNA) in the endosome compartment and its release to the cytosol are the major hurdles for efficient gene transfection. This is generally addressed by using transfection reagents that can overcome these limitations. In this article, the first report is presented which suggests that controlling the release of pDNA from endosome is the key for achieving efficient transfection. In this study, chondroitin sulfate (CS)-coated DNA-nanoplexes are developed using a modular approach where CS is coated post-pDNA/PEI nanoplex formation. To ensure good stability of the nanoplexes, imine/enamine chemistry is exploited by reacting aldehyde-modified chondroitin sulfate (CS-CHO) with free amines of pDNA/PEI complex. This supramolecular nanocarrier system displays efficient cellular uptake, and controlled endosomal pDNA release without eliciting any cytotoxicity. On the contrary, burst release of pDNA from endosome (using chloroqine) results in significant reduction in gene expression. Unlike pDNA/PEI-based transfection, the nanoparticle design presented here shows exceptional stability and gene transfection efficiency in different cell lines such as human colorectal cancer cells (HCT116), human embryonic kidney cells (HEK293), and mouse skin-derived mesenchymal stem cells (MSCs) using luciferase protein as a reporter gene. This new insight will be valuable in designing next generation of transfection reagents.
机译：内体区室中质粒DNA（pDNA）的降解及其向细胞质中的释放是有效基因转染的主要障碍。这通常通过使用可以克服这些限制的转染试剂来解决。在本文中，提出了第一份报告，该报告表明控制内体中pDNA的释放是实现有效转染的关键。在这项研究中，使用模块化方法开发了硫酸软骨素（CS）涂层的DNA纳米复合物，其中CS在pDNA / PEI纳米复合物形成后被涂覆。为确保纳米复合物的良好稳定性，通过使醛修饰的硫酸软骨素（CS-CHO）与pDNA / PEI复合物的游离胺反应来开发亚胺/烯胺化学。该超分子纳米载体系统显示出有效的细胞摄取，并控制了内体pDNA的释放，而没有引起任何细胞毒性。相反，pDNA从内体的突发释放（使用氯奎因）导致基因表达显着降低。与基于pDNA / PEI的转染不同，此处展示的纳米颗粒设计在不同的细胞系（例如人结肠直肠癌细胞（HCT116），人胚胎肾细胞（HEK293）和小鼠皮肤衍生的间充质干细胞）中显示出卓越的稳定性和基因转染效率（MSC）使用萤光素酶蛋白作为报告基因。这一新见解将对设计下一代转染试剂具有重要的意义。

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