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Fabrication of Positively Charged Fluorescent Polymer Nanoparticles for Cell Imaging and Gene Delivery

机译:带正电荷的荧光聚合物纳米粒子的细胞成像和基因传递的制备。

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摘要

Development of efficient non-viral gene delivery vector has aroused great attention in the past few decades. In this study, we reported a new gene delivery vector, positively charged fluorescent conjugated polymer nanoparticles (CPNPs), for efficient gene transfection and in-situ intracellular fluorescence imaging. The microscopic and spectroscopic characterizations demonstrated that these CPNPs possess decent fluorescence performance (e.g. with fluorescence quantum yield of 70.7±0.3%) and small size dimension of ~3.6±0.3 nm (DLS result). Fast and efficient cellular translocation capability was observed according to the time-dependent living cell imaging experiments. Nearly all of the cells were loaded with CPNPs after co-incubation for 2 h regardless of the cell type. In comparison with the commonly used gene delivery vector, lipofectamine 2000 (with gene transfection efficiency of 55±5% for pEGFP), the gene expression efficiency with the positively charged CPNPs (70±3% for pEGFP) was improved significantly. Intracellular fluorescence imaging results demonstrated that the CPNPs could actively assemble close to the periphery of nuclei. Disassembly was not observed even 36 h later, which greatly facilitates releasing of pDNA close to the periphery of nuclei and thus promotes the gene transfection efficiency.
机译:在过去的几十年中,有效的非病毒基因递送载体的开发引起了极大的关注。在这项研究中,我们报告了一种新的基因传递载体,带正电的荧光共轭聚合物纳米粒子(CPNP),用于有效的基因转染和原位细胞内荧光成像。显微和光谱表征表明,这些CPNP具有良好的荧光性能(例如,荧光量子产率为70.7±0.3%),并且小尺寸尺寸为〜3.6±0.3nm(DLS结果)。根据时间依赖性活细胞成像实验观察到快速有效的细胞移位能力。共同孵育2小时后,几乎所有细胞均加载了CPNP,无论细胞类型如何。与常用的基因递送载体lipofectamine 2000(pEGFP的基因转染效率为55±5%)相比,带正电荷的CPNP(pEGFP的基因转染效率为70±3%)显着提高。细胞内荧光成像结果表明,CPNPs可以活跃地靠近细胞核的周围组装。甚至在36小时后仍未观察到拆卸,这极大地促进了pDNA在细胞核外围附近的释放,从而提高了基因转染效率。

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