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Chemical RNA digestion enables robust RNA-binding site mapping at single amino acid resolution

Cited 4 time in Web of Science Cited 3 time in Scopus
Authors
Bae, Jong Woo; Kwon, S. Chul; Na, Yongwoo; Kim, V. Narry; Kim, Jong-Seo
Issue Date
2020-07
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
URI
https://hdl.handle.net/10371/171946
DOI
https://doi.org/10.1038/s41594-020-0436-2
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College of Natural Sciences (자연과학대학)Dept. of Biological Sciences (생명과학부)Journal Papers (저널논문_생명과학부)
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