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Chemical RNA digestion enables robust RNA-binding site mapping at single amino acid resolution
Cited 22 time in
Web of Science
Cited 25 time in Scopus
- Authors
- Issue Date
- 2020-07
- Publisher
- Nature Publishing Group
- Citation
- Nature Structural and Molecular Biology, Vol.27 No.7, pp.678-682
- Abstract
- RNA-binding sites (RBSs) can be identified by liquid chromatography and tandem mass spectrometry analyses of the protein-RNA conjugates created by crosslinking, but RBS mapping remains highly challenging due to the complexity of the formed RNA adducts. Here, we introduce RBS-ID, a method that uses hydrofluoride to fully cleave RNA into mono-nucleosides, thereby minimizing the search space to drastically enhance coverage and to reach single amino acid resolution. Moreover, the simple mono-nucleoside adducts offer a confident and quantitative measure of direct RNA-protein interaction. Using RBS-ID, we profiled similar to 2,000 human RBSs and probedStreptococcus pyogenesCas9 to discover residues important for genome editing.
- ISSN
- 1545-9993
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