Transcriptome-wide quantification of double-stranded RNAs in live mouse tissues by dsRIP-Seq

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).

Automated summary

The presence of dsRNAs in cells plays multiple regulatory functions, with viral dsRNAs activating innate immune responses. Alterations in RNA editing and modifications can lead to accumulation of abnormal endogenous dsRNAs and trigger harmful innate immune responses. This study provides a complete protocol for measuring dsRNAs in live mouse tissue using dsRNA immunoprecipitation and sequencing, focusing on tissue isolation, immunoprecipitation, and computational analysis.