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Ribonucleoprotein Purification And Characterization Using Rna Mango, Shanker Shyam S. Panchapakesan, Matthew L. Ferguson, Eric J. Hayden, Xin Chen, Aaron A. Hoskins, Peter J. Unrau

Physics Faculty Publications and Presentations

The characterization of RNA–protein complexes (RNPs) is a difficult but increasingly important problem in modern biology. By combining the compact RNA Mango aptamer with a fluorogenic thiazole orange desthiobiotin (TO1-Dtb or TO3-Dtb) ligand, we have created an RNA tagging system that simplifies the purification and subsequent characterization of endogenous RNPs. Mango-tagged RNP complexes can be immobilized on a streptavidin solid support and recovered in their native state by the addition of free biotin. Furthermore, Mango-based RNP purification can be adapted to different scales of RNP isolation ranging from pull-down assays to the isolation of large amounts of biochemically defined cellular …