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Cellular dsRNA interactome captured by K1 antibody reveals the regulatory map of exogenous RNA sensing
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Cellular dsRNA interactome captured by K1 antibody reveals the regulatory map of exogenous RNA sensing
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Journal Article
Cellular dsRNA interactome captured by K1 antibody reveals the regulatory map of exogenous RNA sensing
2025
Overview
RNA-binding proteins (RBPs) provide a critical post-transcriptional regulatory layer in determining RNA fate. Currently, UV crosslinking followed by oligo-dT pull-down is the gold standard in identifying the RBP repertoire of poly-adenylated RNAs, but such method is ineffective in capturing RBPs that recognize double-stranded RNAs (dsRNAs). Here, we utilize anti-dsRNA K1 antibody immunoprecipitation followed by quantitative mass spectrometry to comprehensively identify RBPs bound to cellular dsRNAs without external stimulus. Notably, our dsRNA interactome contains proteins involved in sensing N
6
-methyladenosine RNAs and stress granule components. We further perform targeted CRISPR-Cas9 knockout functional screening and discover proteins that can regulate the interferon (IFN) response during exogenous RNA sensing. Interestingly, most dsRBPs promote IFN-β secretion in response to dsRNA stimulation and act as antiviral factors during HCoV-OC43 infection. Our dsRNA interactome capture provides an unbiased and comprehensive characterization of putative dsRBPs and will facilitate our understanding of dsRNA sensing in physiological and pathological contexts.
Exploration of dsRNA interactome using the K1 antibody and functional studies using CRISPR-Cas9 knockout screening of these dsRBP candidates elucidate the role of dsRNA interactome in immune responses and their importance in viral infection.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
