Wakimoto Hiroko, Seidman J G, Foo Roger S Y, Jiang Jianming
Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Cardiovascular Research Institute (CVRI), Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Curr Protoc Mol Biol. 2016 Jul 1;115:23.16.1-23.16.9. doi: 10.1002/cpmb.9.
RNA interference (RNAi) is a rapid approach to dissect loss-of-function phenotype for a gene of interest. However, it is challenging to perform RNAi in specific organs and tissues in vivo. Engineered viruses can provide a useful tool for delivery of small RNAs in vivo. Recombinant adeno-associated viruses (rAAVs) are the preferred method for delivering genes or gene modulators to target cells due to their high titer, low immune response, ability to transduce many types of cell, and overall safety. In this unit, we describe protocols for use of rAAVs as a cargo to deliver miRNA backbone-based shRNA controlled by a cardiac-specific promoter into the mouse heart. © 2016 by John Wiley & Sons, Inc.
RNA干扰（RNAi）是一种剖析感兴趣基因功能缺失表型的快速方法。然而，在体内特定器官和组织中进行RNAi具有挑战性。工程病毒可为体内小RNA的递送提供有用工具。重组腺相关病毒（rAAV）因其高滴度、低免疫反应、能够转导多种类型细胞以及整体安全性，是将基因或基因调节剂递送至靶细胞的首选方法。在本单元中，我们描述了使用rAAV作为载体，将由心脏特异性启动子控制的基于miRNA骨架的短发夹RNA（shRNA）递送至小鼠心脏的实验方案。© 2016约翰威立国际出版公司。
