通过脾内注射血清型8腺相关病毒(AAV8)递送的小发夹RNA在肝脏中高效敲除基因

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES
Xiaoqin Liu, Robert Y L Tsai
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引用次数: 0

摘要

肝脏是执行重要代谢功能的主要器官。开发一种高效安全的方法来敲除肝脏中的基因表达,为确定肝脏病理生理学中的基因功能提供了重要工具。在这项研究中,我们介绍了一种脾内注射血清型8腺相关病毒(AAV8)的方法,AAV8可表达针对感兴趣的靶基因--核固蛋白(NS)的小发夹RNA(shRNA)。脾内注射表达NS靶向shRNA的AAV8(AAV8-shNS1)与门静脉注射表达乱序shRNA的AAV8(AAV8-shScr)相比,肝脏中NS的敲除效率相同。此外,注射AAV8-shRNA病毒在肝实质中引发的炎症反应极小。最重要的是,这种脾内注射方案对技术要求不高,而且注射部位出血量极少,而这正是门静脉注射导致围手术期和术后死亡的主要原因。这项研究报告了一种在肝脏中实现高效基因敲除的改良且相对安全的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Efficient Gene Knockdown in the Liver via Intrasplenic Injection of Adeno-Associated Virus Serotype 8 (AAV8)-Delivered Small Hairpin RNA.

The liver is a major organ that performs essential metabolic functions. Developing an efficient and safe method to knock down gene expression in the liver provides an important tool for determining gene function in liver pathophysiology. In this study, we describe a method for intrasplenic injection of adeno-associated virus serotype 8 (AAV8) engineered to express a small hairpin RNA (shRNA) against a target gene of interest, nucleostemin (NS). Intrasplenic injection of AAV8 expressing an NS-targeting shRNA (AAV8-shNS1) achieved the same knockdown efficiency of NS in the liver as did portal vein injection, compared to the injection of AAV8 expressing a scrambled sequence shRNA (AAV8-shScr). Furthermore, injection of the AAV8-shRNA virus triggered minimal inflammatory reactions in the liver parenchyma. Most importantly, this intrasplenic injection protocol was not technically demanding and caused minimal bleeding at the injection site, which is the leading cause of perioperative and postoperative mortality when performing portal vein injection. This study reports an improved and relatively safe method to achieve efficient gene knockdown in the liver.

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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
期刊介绍: JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.
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