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首页> 外文期刊>Ultrasound in Medicine and Biology >SONOPORATION AS AN APPROACH FOR SIRNA DELIVERY INTO T CELLS
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SONOPORATION AS AN APPROACH FOR SIRNA DELIVERY INTO T CELLS

机译：Sonoporation作为siRNA递送到T细胞的方法

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Delivery of small interfering RNAs (siRNAs) into primary T cells is quite challenging because they are non-proliferating cells and are difficult to transfect with non-viral approaches. Because sonoporation is independent of the proliferation status of cells and siRNA acts in the cell cytoplasm, we investigated whether sonoporation could be used to deliver siRNA into mouse and human T cells. Cells mixed with Definity microbubbles and siRNA were sonicated with a non-focused transducer of center frequency 2.20 MHz producing ultrasound at a 10% duty cycle, pulse repetition frequency of 2.20 kHz and spatial average temporal average ultrasound intensity of 1.29 W/cm(2) for 5 s and then examined for siRNA fluorescence by flow cytometry analysis. These sonoporation conditions resulted in high-efficiency transfection of siRNA in mouse and human T cells. Further, the efficacy of siRNA delivery by sonoporation was illustrated by the successful visualization of decreased methylation-controlled J protein expression in mouse and human CD8 T cells via Western blot analysis. The results provide the first evidence that sonoporation is a novel approach to delivery of siRNA into fresh isolated mouse and human T cells in vitro, and might be used for in vivo studies in the future. (C) 2019 Published by Elsevier Inc. on behalf of World Federation for Ultrasound in Medicine & Biology.
机译：将小干扰RNA（siRNA）分泌到原发性T细胞中是非常具有挑战性的，因为它们是未增殖的细胞，并且难以通过非病毒方法转染。因为声孔与细胞和siRNA的增殖状态无关，所以我们研究了声孔是否可用于将siRNA递送到小鼠和人体T细胞中。用明确微泡和siRNA混合的细胞用420MHz的非聚焦换能器以10％占空比产生超声波的非聚焦传感器，脉冲重复频率为2.20kHz，空间平均时间平均超声强度为1.29 w / cm（2）通过流式细胞术分析检查5秒，然后检查siRNA荧光。这些声孔条件导致小鼠和人T细胞中siRNA的高效转染。此外，通过蛋白质印迹分析的成功可视化通过蛋白质印迹分析成功可视化通过蛋白质印迹和人CD8T细胞中的甲基化控制的J蛋白表达的成功可视化来说明SiRNA递送的疗效。结果提供了第一种证据表明Sonoporation是一种新的方法，将siRNA递送到鲜物孤立的小鼠和人T细胞中，并且可能用于将来的体内研究。 （c）2019年由elsevier Inc.发布的，代表世界医学与生物学的超声联联联合会。

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