Nonviral ultrasound-mediated gene delivery in small and large animal models
Nonviral ultrasound-mediated gene delivery in small and large animal models
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DOI：
10.1038/s41596-019-0125-y
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发表时间：
2019-04-01
期刊：
影响因子：
14.8
通讯作者：
Gazit, Dan
中科院分区：
文献类型：
--
作者：
Bez, Maxim;Foiret, Josquin;Gazit, Dan
超声介导的基因治疗是一种微创、无病毒、临床可翻译的基因治疗方法。与病毒载体相比，这种方法具有良好的安全性，与其他物理基因传递方法(如电穿孔)相比，侵入性更小。我们以前曾使用超声修复在不同的骨骼组织中过表达转基因，以诱导组织再生。在这里，我们提供了一种可以很容易地修改的方案，以解决各种其他组织再生或其他应用的目标，例如癌症和神经退行性疾病。该方案描述了如何在小鼠和猪的动物模型中准备、引导和优化超声介导的基因传递。该方案包括制备微泡-DNA混合物和在超声成像下的体内超声手术。超声介导的基因传递可以在10分钟内完成。在DNA传递后，动物可以被跟踪监测基因表达、蛋白质分泌和其他转基因特有的结果，包括组织再生。这一过程可以由有超声成像或活体操作经验的有能力的研究生或技术人员完成。
Ultrasound-mediated gene delivery (sonoporation) is a minimally invasive, nonviral and clinically translatable method of gene therapy. This method offers a favorable safety profile over that of viral vectors and is less invasive as compared with other physical gene delivery approaches (e.g., electroporation). We have previously used sonoporation to overexpress transgenes in different skeletal tissues in order to induce tissue regeneration. Here, we provide a protocol that could easily be adapted to address various other targets of tissue regeneration or additional applications, such as cancer and neurodegenerative diseases. This protocol describes how to prepare, conduct and optimize ultrasound-mediated gene delivery in both a murine and a porcine animal model. The protocol includes the preparation of a microbubble-DNA mix and in vivo sonoporation under ultrasound imaging. Ultrasound-mediated gene delivery can be accomplished within 10 min. After DNA delivery, animals can be followed to monitor gene expression, protein secretion and other transgene-specific outcomes, including tissue regeneration. This procedure can be accomplished by a competent graduate student or technician with prior experience in ultrasound imaging or in performing in vivo procedures.