RNA‐Cleaving Deoxyribozymes Differentiate Methylated Cytidine Isomers in RNA. Issue 35 (26th July 2021)

• Record Type: Journal Article
• Authors: Liaqat, Anam; Sednev, Maksim V.; Stiller, Carina; Höbartner, Claudia
• Is Part Of: Angewandte Chemie international edition. Volume 60:Issue 35(2021)
• Page Start: 19058
• Page End: 19062
• Publication Date: 2021-07-26
• Subjects: deoxyribozymes -- epitranscriptomics -- in vitro selection -- RNA modification -- site-specific RNA cleavage; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.202106517
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Deoxyribozymes are emerging as modification‐specific endonucleases for the analysis of epigenetic RNA modifications. Here, we report RNA‐cleaving deoxyribozymes that differentially respond to the presence of natural methylated cytidines, 3‐methylcytidine (m 3 C), N 4 ‐methylcytidine (m 4 C), and 5‐methylcytidine (m 5 C), respectively. Using in vitro selection, we found several DNA catalysts, which are selectively activated by only one of the three cytidine isomers, and display 10‐ to 30‐fold accelerated cleavage of their target m 3 C‐, m 4 C‐ or m 5 C‐modified RNA. An additional deoxyribozyme is strongly inhibited by any of the three methylcytidines, but effectively cleaves unmodified RNA. The m X C‐detecting deoxyribozymes are programmable for the interrogation of natural RNAs of interest, as demonstrated for human mitochondrial tRNAs containing known m 3 C and m 5 C sites. The results underline the potential of synthetic functional DNA to shape highly selective active sites. Abstract : In vitro selected deoxyribozymes respond to the presence of methylcytidines m 3 C, m 4 C and m 5 C in RNA with either enhanced or strongly diminished cleavage rates, and thereby directly reveal the presence of the modified cytidines in target RNA.