Chemical Method to Sequence 5-Formylcytosine on RNA.

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 (f⁵C) modifications on RNA based on the selective and efficient malononitrile-mediated labeling of f⁵C 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 f⁵C at the wobble position of mt-tRNA(Met) in different organisms and tissue types and find that high-level f⁵C modification is present in mammals but lacking in lower eukaryotes. Our work sheds light

SUBMITTER: Li A 

PROVIDER: S-EPMC9357364 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature