Wei Lin, Zhang Di, Zheng Xuanfang, Zeng Xuyao, Zeng Youlin, Shi Xinbo, Su Xin, Xiao Lehui
Key Laboratory of Chemical Biology & Traditional Chinese Medicine Research, Ministry of Education, Key Laboratory of Phytochemical R&D of Hunan Province, College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, Hunan, 410081, China.
State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, College of Chemistry, Nankai University, Tianjin, 300071, China.
Nanotheranostics. 2018 Feb 15;2(2):157-167. doi: 10.7150/ntno.22988. eCollection 2018.
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.
在过去几十年中,高效非病毒基因递送载体的研发引起了广泛关注。在本研究中,我们报道了一种新型基因递送载体——带正电荷的荧光共轭聚合物纳米颗粒(CPNPs),用于高效基因转染和原位细胞内荧光成像。显微镜和光谱表征表明,这些CPNPs具有良好的荧光性能(例如荧光量子产率为70.7±0.3%),且尺寸较小,约为3.6±0.3 nm(动态光散射结果)。根据时间依赖性活细胞成像实验,观察到其具有快速高效的细胞转运能力。无论细胞类型如何,共孵育2小时后几乎所有细胞都被CPNPs负载。与常用的基因递送载体Lipofectamine 2000(对pEGFP的基因转染效率为55±5%)相比,带正电荷的CPNPs对pEGFP的基因表达效率(70±3%)显著提高。细胞内荧光成像结果表明,CPNPs能够在细胞核周边主动聚集。即使36小时后也未观察到解聚现象,这极大地促进了pDNA在细胞核周边的释放,从而提高了基因转染效率。