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A chemical method to sequence 5-formylcytosine on RNA

Ang Li, Xuemeng Sun, A. Emilia Arguello, Ralph E. Kleiner
doi: https://doi.org/10.1101/2021.11.23.469780
Ang Li
Department of Chemistry, Princeton University, Princeton, NJ 08544
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Xuemeng Sun
Department of Chemistry, Princeton University, Princeton, NJ 08544
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A. Emilia Arguello
Department of Chemistry, Princeton University, Princeton, NJ 08544
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Ralph E. Kleiner
Department of Chemistry, Princeton University, Princeton, NJ 08544
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  • For correspondence: rkleiner@princeton.edu
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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 to sequence 5-formylcytosine (f5C) modifications on RNA based upon selective and efficient malononitrile-mediated labeling of f5C residues to generate adducts that are read as C-to-T mutations upon reverse transcription and PCR 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 on mitochondrial tRNA modifications throughout eukaryotic evolution and provides a general platform for characterizing the f5C epitranscriptome.

Competing Interest Statement

The authors have declared no competing interest.

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Posted November 23, 2021.
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A chemical method to sequence 5-formylcytosine on RNA
Ang Li, Xuemeng Sun, A. Emilia Arguello, Ralph E. Kleiner
bioRxiv 2021.11.23.469780; doi: https://doi.org/10.1101/2021.11.23.469780
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A chemical method to sequence 5-formylcytosine on RNA
Ang Li, Xuemeng Sun, A. Emilia Arguello, Ralph E. Kleiner
bioRxiv 2021.11.23.469780; doi: https://doi.org/10.1101/2021.11.23.469780

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