Precision-Guided Nanospears for Targeted and High-Throughput Intracellular Gene Delivery
Precision-Guided Nanospears for Targeted and High-Throughput Intracellular Gene Delivery
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用于靶向和高通量细胞内基因传递的精确引导纳米矛
DOI：
10.1021/acsnano.8b00763
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发表时间：
2018
期刊：
影响因子：
17.1
通讯作者：
Weiss Paul S.
中科院分区：
文献类型：
--
作者：
Xu Xiaobin;Hou Shuang;Wattanatorn Natcha;Wang Fang;Yang Qin;Zhao Chuanzhen;Yu Xiao;Tseng Hsian-Rong;Jonas Steven J.;Weiss Paul S.
报道了一种基于磁性纳米矛的高效非病毒平台，用于高通量和亚细胞精确靶向细胞内核酸递送。这些磁性纳米矛由Au/Ni/Si（长度约5 μm，尖端直径<50 nm）制成，并通过纳米球光刻和金属沉积制备。磁铁用于指导单个纳米矛的机械运动，从而实现精确控制位置和三维旋转。这些纳米矛在从硅衬底释放之前，通过一层一层的方法，用增强的绿色荧光蛋白（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接近目标粘附的U87胶质母细胞瘤细胞，穿透细胞膜，使质粒货物在细胞内递送。24 h后，靶细胞发出绿色荧光，表明转染成功。这种纳米矛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。细胞存活率>90%，转染率>80%，超过了传统的非病毒细胞内方法。这种方法与良好生产规范相兼容，规避了新兴细胞疗法转化和临床部署的障碍。
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.