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首页> 外文期刊>Ultrasound in Medicine and Biology >Shock waves and DNA-cationic lipid assemblies: A synergistic approach to express exogenous genes in human cells
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Shock waves and DNA-cationic lipid assemblies: A synergistic approach to express exogenous genes in human cells

机译:冲击波和DNA阳离子脂质组装体:在人类细胞中表达外源基因的协同方法

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Cationic lipid/DNA complexes (lipoplexes) represent a powerful tool for cell transfection; however, their use is still limited by important concerns, including toxicity and poor internalization into deep tissues. In this work, we investigated the use of shock wave-induced acoustic cavitation invitro for the transfection of lipoplexes in human embryo kidney 293cells. We selected shock waves with the ability to internalize 10-kDa fluorescein isothiocyanate-dextran into cells while maintaining survival rates above 50%. Cell transfection was tested using the green fluorescent protein-encoding plasmid pCX::GFPGPI2. Confocal microscopy and fluorescence-assisted cell sorting analyses revealed successful transfection after treatments ranging from 1 to 3min using 60 to 180 shock waves at peak amplitudes of 12.3±1.5MPa. Interestingly, the combination of shock waves and lipoplexes induced a 3.1- and 3.8-fold increase in the expression of the reporter gene compared with the use of lipoplexes or shock waves alone, respectively. These results indicate that cationic DNA assembly and shock waves act in a synergistic manner to promote transfection of human cells, revealing a potential approach for non-invasive site-specific gene therapy.
机译:阳离子脂质/ DNA复合物(脂质复合物)是细胞转染的有力工具。然而,它们的使用仍然受到重要问题的限制,包括毒性和深层组织内化不良。在这项工作中,我们研究了使用冲击波诱导的声空化体外转染人胚肾293细胞中的脂质复合体。我们选择了能够将10 kDa异硫氰酸荧光素-葡聚糖内化到细胞中,同时将存活率保持在50%以上的冲击波。使用绿色荧光蛋白编码质粒pCX :: GFPGPI2测试细胞转染。共聚焦显微镜和荧光辅助细胞分选分析显示,在60分钟至180分钟的冲击波下,以12.3±1.5MPa的峰值振幅处理1至3分钟后,成功转染。有趣的是,与单独使用脂质体或冲击波相比,冲击波和脂质体的组合分别引起报告基因表达的3.1倍和3.8倍增加。这些结果表明阳离子DNA组装和冲击波以协同方式促进人类细胞的转染,从而揭示了非侵入性位点特异性基因治疗的潜在方法。

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