Chemical Method to Sequence 5-Formylcytosine on RNA. Author Ang Li, Xuemeng Sun, A Emilia Arguello, Ralph Kleiner Publication Year 2022 Type Journal Article Abstract Epitranscriptomic RNA modifications can regulate biological processes, but there remains a major gap in our ability to identify and measure individual modifications at nucleotide resolution. Here we present Mal-Seq, a chemical method for sequencing 5-formylcytosine (fC) modifications on RNA based on the selective and efficient malononitrile-mediated labeling of fC residues to generate adducts that are read as C-to-T mutations upon reverse transcription and polymerase chain reaction amplification. We apply Mal-Seq to characterize the prevalence of fC at the wobble position of mt-tRNA(Met) in different organisms and tissue types and find that high-level fC modification is present in mammals but lacking in lower eukaryotes. Our work sheds light on mitochondrial tRNA modifications throughout eukaryotic evolution and provides a general platform for characterizing the fC epitranscriptome. Keywords Animals, RNA Processing, Post-Transcriptional, RNA, Mammals, RNA, Transfer, Cytosine Journal ACS Chem Biol Volume 17 Issue 3 Pages 503-508 Date Published 2022 Mar 18 ISSN Number 1554-8937 DOI 10.1021/acschembio.1c00707 Alternate Journal ACS Chem Biol PMCID PMC9357364 PMID 35212224 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML