Systematic and Comprehensive Analysis of Interferon Response induced by siRNA

siRNA 诱导的干扰素反应的系统综合分析

• 批准号: 16300151
• 负责人: MIYAGISHI Makoto
• 金额: $ 9.15万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16300151/
• 关键词: Interferon Response; RNA interference; Biotechnology; siRNA; ジーンディスカバリー; アポトシス; RNAiライブラリー

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.

在哺乳动物细胞中，siRNA已用于诱导RNA干扰（RNAi），以试图阻止由大于30 bp的长双链RNA（dsRNA）引起的非特异性效应（包括干扰素（IFN）应答）。在本项目中，我们开发了一种新的和简单的策略，以避免激活的干扰素反应的dsRNA，并分析干扰素通路的siRNA文库的手段。我们发现，超过100 bp的修饰的发夹RNA（mhRNA），在正义链内具有多个特定的点突变，并从U6或tRNAVa 1启动子转录，可以引起RNAi而不诱导IFN途径基因。这些发现将促进RNAi在哺乳动物细胞中的开发，特别是在针对病原性病毒的RNAi治疗领域。siRNA文库筛选结果显示，dsRNA诱导的细胞凋亡途径包括JNK/SAPK介导的线粒体途径和ERK 2相关途径。

项目成果

Intracellular-diced dsRNA has enhanced efficacy for silencing HCV RNA and overcomes variation in the viral genotype

• DOI: 10.1038/sj.gt.3302734
• 发表时间: 2006-06-01
• 期刊: GENE THERAPY
• 影响因子: 5.1
• 作者: Watanabe, T.;Sudoh, M.;Kohara, M.
• 通讯作者: Kohara, M.

Analysis of double-stranded RNA-induced apoptosis pathways using interferon-response noninducible small interfering RNA expression vector library

• DOI: 10.1074/jbc.m412784200
• 发表时间: 2005-07-08
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Matsumoto, S;Miyagishi, M;Taira, K
• 通讯作者: Taira, K

Gene therapy for human small-cell lung carcinoma by inactivation of Skp-2 with virally mediated RNA interference

• DOI: 10.1038/sj.gt.3302391
• 发表时间: 2005-01-01
• 期刊: GENE THERAPY
• 影响因子: 5.1
• 作者: Sumimoto, H;Yamagata, S;Kawakami, Y
• 通讯作者: Kawakami, Y

The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses

• DOI: 10.1038/ni1087
• 发表时间: 2004-07-01
• 期刊: NATURE IMMUNOLOGY
• 影响因子: 30.5
• 作者: Yoneyama, M;Kikuchi, M;Fujita, T
• 通讯作者: Fujita, T

Generation of shRNAi expression library against the whole human transcripts.

针对整个人类转录本生成 shRNAi 表达文库。

• 发表时间: 2004
• 期刊: Virus Res. 102
• 作者: Miyagishi;M.;Matsumoto;S.;Taira;K.
• 通讯作者: K.