Chalupnikova Katerina, Nejepinska Jana, Svoboda Petr
Institute of Molecular Genetics AS CR, Prague, Czech Republic.
Methods Mol Biol. 2013;942:291-314. doi: 10.1007/978-1-62703-119-6_16.
Double-stranded RNA (dsRNA) is involved in different biological processes. At least three different pathways can respond to dsRNA in mammals. One of these pathways is RNA interference (RNAi) where long dsRNA induces sequence-specific degradation of transcripts carrying sequences complementary to dsRNA. Long dsRNA is also a potent trigger of the interferon pathway, a sequence-independent response that leads to global suppression of translation and global RNA degradation. In addition, dsRNA can be edited by adenosine deamination, which may result in nuclear retention and degradation of dsRNA or in alteration of RNA coding potential. Here, we provide a technical review summarizing different strategies of long dsRNA usage. While the review is largely focused on long dsRNA-induced RNAi in mammalian cells, it also provides helpful information on both the in vitro production and in vivo expression of dsRNAs. We present an overview of currently available vectors for dsRNA expression and provide the latest update on oocyte-specific transgenic RNAi approaches.
双链RNA（dsRNA）参与不同的生物学过程。在哺乳动物中，至少有三种不同的途径可对dsRNA作出反应。其中一种途径是RNA干扰（RNAi），即长双链RNA诱导携带与dsRNA互补序列的转录本发生序列特异性降解。长双链RNA也是干扰素途径的有效触发因素，这是一种不依赖序列的反应，会导致整体翻译抑制和整体RNA降解。此外，dsRNA可通过腺苷脱氨作用进行编辑，这可能导致dsRNA的核滞留和降解，或导致RNA编码潜能的改变。在此，我们提供一篇技术综述，总结长双链RNA使用的不同策略。虽然该综述主要关注哺乳动物细胞中长双链RNA诱导的RNAi，但它也提供了有关dsRNA体外生产和体内表达的有用信息。我们概述了目前可用于dsRNA表达的载体，并提供了卵母细胞特异性转基因RNAi方法的最新进展。
