ICLAMP: a novel technique to explore adenosine deamination via inosine chemical labeling and affinity molecular purification.

Autor: Yang Y; Research Institute for Biomedical Sciences, Tokyo University of Science, Chiba, Japan., Nakayama K; Research Institute for Biomedical Sciences, Tokyo University of Science, Chiba, Japan., Okada S; Department of Microbiology, Faculty of Medicine, Shimane University, Izumo-shi, Japan., Sato K; Department of Medicinal and Life Sciences, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda-shi, Japan., Wada T; Department of Medicinal and Life Sciences, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda-shi, Japan., Sakaguchi Y; Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Bunkyo-ku, Japan., Murayama A; Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Bunkyo-ku, Japan., Suzuki T; Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Bunkyo-ku, Japan., Sakurai M; Research Institute for Biomedical Sciences, Tokyo University of Science, Chiba, Japan.
Jazyk: angličtina
Zdroj: FEBS letters [FEBS Lett] 2024 May; Vol. 598 (9), pp. 1080-1093. Date of Electronic Publication: 2024 Mar 24.
DOI: 10.1002/1873-3468.14854
Abstrakt: Recent developments in sequencing and bioinformatics have advanced our understanding of adenosine-to-inosine (A-to-I) RNA editing. Surprisingly, recent analyses have revealed the capability of adenosine deaminase acting on RNA (ADAR) to edit DNA:RNA hybrid strands. However, edited inosines in DNA remain largely unexplored. A precise biochemical method could help uncover these potentially rare DNA editing sites. We explore maleimide as a scaffold for inosine labeling. With fluorophore-conjugated maleimide, we were able to label inosine in RNA or DNA. Moreover, with biotin-conjugated maleimide, we purified RNA and DNA containing inosine. Our novel technique of inosine chemical labeling and affinity molecular purification offers substantial advantages and provides a versatile platform for further discovery of A-to-I editing sites in RNA and DNA.
(© 2024 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)
Databáze: MEDLINE
Externí odkaz: