Systematic and Comprehensive Analysis of Interferon Response induced by siRNA

在哺乳动物细胞中，sirna已被用于诱导RNA干扰（RNAi），以防止由超过30 bp的长双链RNA （dsRNAs）引起的非特异性效应（包括干扰素（IFN）反应）。在本项目中，我们开发了一种新颖而简单的策略来避免dsRNA激活IFN反应，并通过siRNA文库分析干扰素途径。我们发现，超过100 bp的修饰发夹rna (mhRNAs)，在感觉链内具有多个特定的点突变，并从U6或tRNAVa1启动子转录，可以在不诱导IFN通路基因的情况下引起RNAi。这些发现将加强RNAi在哺乳动物细胞中的应用，特别是在RNAi治疗致病性病毒领域。此外，siRNA文库筛选结果显示，dsRNA诱导的凋亡途径包括JNK/ sapk介导的线粒体途径和erk2相关途径。

In mammalian cells, siRNAs have been used to induce RNA interference (RNAi) in an attempt to prevent nonspecific effects (including the interferon (IFN) response) which are caused by long double-stranded RNAs (dsRNAs) of more than 30 bp. In this project, we developed a novel and simple strategy for avoiding activation of the IFN response by dsRNA, and analyzed interferon pathway by means of siRNA library. We showed that modified hairpin-RNAs (mhRNAs) of more than 100 bp, with multiple specific point-mutations within the sense strand and transcribed from the U6 or tRNAVa1 promoters, can cause RNAi without inducing the IFN pathway genes. These findings should enhance the exploitation of RNAi in mammalian cells, especially in the field of RNAi therapy against pathogenic viruses. Furthermore the results from screening by siRNA library showed that apoptosis pathway induced by dsRNA include a JNK/SAPK-mediated mitochondrial pathway and an ERK2-related pathway.