Section of Hematology, Yale Cancer Center and Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520, USA.
Yale Stem Cell Center and Yale RNA Center, Yale University School of Medicine, New Haven, CT 06520, USA.
STAR Protoc. 2021 Mar 18;2(1):100366. doi: 10.1016/j.xpro.2021.100366. eCollection 2021 Mar 19.
Double-stranded RNAs (dsRNAs) are abundantly present in cells, playing multiple regulatory functions. dsRNAs of viral origin activate innate immune responses. Since RNA editing and modifications affect the structure and recognition of RNAs, their alteration can result in the accumulation of aberrant endogenous dsRNAs inducing a deleterious innate immune response. Here, we present a complete protocol for the measurement of dsRNAs in a live mouse tissue using dsRNA immunoprecipitation and sequencing (dsRIP-Seq). This protocol focuses on tissue isolation, dsRNA immunoprecipitation and downstream computational analysis. For complete details on the use and execution of this protocol, please refer to Gao et al. (2020).
双链 RNA（dsRNAs）在细胞中大量存在，发挥多种调节功能。病毒来源的 dsRNAs 激活先天免疫反应。由于 RNA 编辑和修饰会影响 RNA 的结构和识别，它们的改变可能导致异常内源性 dsRNAs 的积累，从而诱导有害的先天免疫反应。在这里，我们提供了一种使用双链 RNA 免疫沉淀和测序（dsRIP-Seq）在活体小鼠组织中测量 dsRNAs 的完整方案。该方案侧重于组织分离、dsRNA 免疫沉淀和下游计算分析。有关该方案的使用和执行的完整详细信息，请参阅 Gao 等人。（2020 年）。
