ChIRP-MS: RNA-Directed Proteomic Discovery
Comprehensive identification of RNA-binding proteins by mass spectrometry (ChIRP-ms) is a novel technique for studying endogenous ribonucleoprotein complexes. ChIRP-ms is robust across a wide range of expression level, from abundant housekeeping RNAs (e.g., spliceosomal U RNAs) to relatively lowly expressed RNAs (e.g., Xist). In vivo RNA–protein interactions are chemically cross-linked, and purified using biotinylated antisense oligonucleotides against RNA of interest. Coprecipitated proteins are gently eluted, and identified by mass-spectrometry (for discovery) or by western blotting (for validation).
Key wordslncRNA RNA-binding proteins Mass-spec
This work was supported by the Singapore Agency for Science, Technology, and Research (A*STAR to C.C.) and the US National Institutes of Health and Howard Hughes Medical Institute. (H.Y.C.).
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