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Precision-Guided Nanospears for Targeted and High-Throughput Intracellular Gene Delivery
ACS Nano ( IF 17.1 ) Pub Date : 2018-03-14 00:00:00 , DOI: 10.1021/acsnano.8b00763
Xiaobin Xu 1, 2 , Shuang Hou 1, 3 , Natcha Wattanatorn 1, 2 , Fang Wang 1, 3, 4 , Qing Yang 1, 2 , Chuanzhen Zhao 1, 2 , Xiao Yu 1, 3 , Hsian-Rong Tseng 1, 3 , Steven J Jonas 1, 5, 6, 7 , Paul S Weiss 1, 2, 8
Affiliation  

An efficient nonviral platform for high-throughput and subcellular precision targeted intracellular delivery of nucleic acids in cell culture based on magnetic nanospears is reported. These magnetic nanospears are made of Au/Ni/Si (∼5 μm in length with tip diameters <50 nm) and fabricated by nanosphere lithography and metal deposition. A magnet is used to direct the mechanical motion of a single nanospear, enabling precise control of position and three-dimensional rotation. These nanospears were further functionalized with enhanced green fluorescent protein (eGFP)-expression plasmids via a layer-by-layer approach before release from the underlying silicon substrate. Plasmid functionalized nanospears are guided magnetically to approach target adherent U87 glioblastoma cells, penetrating the cell membrane to enable intracellular delivery of the plasmid cargo. After 24 h, the target cell expresses green fluorescence indicating successful transfection. This nanospear-mediated transfection is readily scalable for the simultaneous manipulation of multiple cells using a rotating magnet. Cell viability >90% and transfection rates >80% were achieved, which exceed conventional nonviral intracellular methods. This approach is compatible with good manufacturing practices, circumventing barriers to the translation and clinical deployment of emerging cellular therapies.

中文翻译:

用于靶向和高通量细胞内基因传递的精确制导纳米矛

报道了一种基于磁性纳米矛的高效非病毒平台,用于在细胞培养物中高通量和亚细胞精确靶向细胞内递送核酸。这些磁性纳米矛由 Au/Ni/Si(长约 5 μm,尖端直径 < 50 nm)制成,并通过纳米球光刻和金属沉积制造。磁铁用于引导单个纳米矛的机械运动,从而能够精确控制位置和三维旋转。这些纳米矛在从下面的硅基板上释放之前,通过逐层方法用增强型绿色荧光蛋白 (eGFP) 表达质粒进一步功能化。质粒功能化的纳米矛被磁性引导以接近目标粘附的 U87 胶质母细胞瘤细胞,穿透细胞膜以实现质粒货物的细胞内递送。24小时后,靶细胞发出绿色荧光,表明转染成功。这种 nanospear 介导的转染很容易扩展,可以使用旋转磁铁同时操作多个细胞。实现了 >90% 的细胞活力和 >80% 的转染率,这超过了传统的非病毒细胞内方法。这种方法符合良好的制造规范,规避了新兴细胞疗法的转化和临床部署障碍。实现了 90% 和 >80% 的转染率,这超过了传统的非病毒细胞内方法。这种方法符合良好的制造规范,规避了新兴细胞疗法的转化和临床部署障碍。实现了 90% 和 >80% 的转染率,这超过了传统的非病毒细胞内方法。这种方法符合良好的制造规范,规避了新兴细胞疗法的转化和临床部署障碍。
更新日期:2018-03-14
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