Isolation of RIG-I-associated RNAs from virus-infected cells.
Isolation of RIG-I-associated RNAs from virus-infected cells.
复制标题
从病毒感染的细胞中分离 RIG-I 相关 RNA
DOI：
10.1007/978-1-4939-0882-0_4
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发表时间：
2014
影响因子：
--
通讯作者：
Rothenfusser
中科院分区：
文献类型：
--
作者：
Schmidt;Linder;Linder;Rothenfusser
当一种新的先天模式识别受体（PRR）被发现时，一个问题立即出现：它能识别哪种分子模式？可以用来回答核酸结合受体的这个问题的一种方法是对合成配体（DNA、RNA或杂交体）进行详细分析，以缩小激活给定受体的最小模式。然而，这并不总是会带来令人满意的答案。解决这个问题的一个补充方法，尽管技术上要求更高，是检查在细胞感染的情况下哪些核酸实际上与受体结合。在这里，我们描述了一个基本的协议，以分离RNA结合到RNA受体的RIG-I样解旋酶家族从病毒感染的细胞通过免疫沉淀（IP）。然后，分离的RNA可以用于用各种方法分析其来源、特征和免疫刺激特性。
When a novel innate pattern recognition receptor (PRR) is identified, a question comes up immediately: Which molecular pattern(s) can it recognize? One approach that can be taken to answer this question for nucleic acid-binding receptors is the detailed analysis of synthetic ligands (DNA, RNA, or hybrids) to narrow in on the minimal patterns that activate a given receptor. However, this may not always lead to a satisfying answer. A complementary albeit technically more demanding way to tackle this question is to examine which nucleic acids are actually bound by the receptor in a setting of cellular infection. Here, we describe a basic protocol to isolate RNAs bound to the RNA receptors of the RIG-I-like helicase family from virus-infected cells via immunoprecipitation (IP). The isolated RNA can then be used to analyze its origin, characteristics, and immunostimulatory properties with a variety of methods.
影响因子：
32.4
作者：
Schlee, Martin;Roth, Andreas;Hornung, Veit;Hagmann, Cristina Amparo;Wimmenauer, Vera;Barchet, Winfried;Coch, Christoph;Janke, Markus;Mihailovic, Aleksandra;Wardle, Greg;Juranek, Stefan;Kato, Hiroki;Kawai, Taro;Poeck, Hendrik;Fitzgerald, Katherine A.;Takeuchi, Osamu;Akira, Shizuo;Tuschl, Thomas;Latz, Eicke;Ludwig, Janos;Hartmann, Gunther
通讯作者：
Hartmann, Gunther