Abstract
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).
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Acknowledgments
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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Chu, C., Chang, H.Y. (2018). ChIRP-MS: RNA-Directed Proteomic Discovery. In: Sado, T. (eds) X-Chromosome Inactivation. Methods in Molecular Biology, vol 1861. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8766-5_3
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DOI: https://doi.org/10.1007/978-1-4939-8766-5_3
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