@misc{hartstock_ovcharenko_kueck_spacek_cornelissen_huewel_dieterich_rentmeister_2022,
title={MePMe-seq: Antibody-free simultaneous m6A and m5C mapping in mRNA by metabolic propargyl labeling and sequencing},
DOI={10.1101/2022.03.16.484494},
abstractNote={Internal modifications of mRNA have emerged as widespread and versatile regulatory mechanism to control gene expression at the post-transcriptional level. Current insights rely on the ability to make a modified nucleoside amenable to sequencing. Most of the modifications are methylations involving the co-factor S-adenosyl-L-methionine (SAM), however, simultaneous detection of different methylation sites in the same sample has remained elusive. We present metabolic labeling with propargyl-selenohomocysteine (PSH) in combination with click chemistry to detect N6-methyladenosine (m6A) and 5-methylcytidine (m5C) sites in mRNA with single nucleotide precision in the same sequencing run (MePMe-seq). Intracellular formation of the corresponding SAM analogue leads to detectable levels of N6-propargyl-A (prop6A) and 5-propargyl-C (prop5C). MePMe-seq overcomes the problems of antibodies for enrichment and sequence-motifs for evaluation, limiting previous methodologies. The joint evaluation of m6A and m5C sites opens the door to study their interconnectivity and improve our understanding of mechanisms and functions of the RNA methylome.},
publisher={Cold Spring Harbor Laboratory},
author={Hartstock, Katja and Ovcharenko, Anna and Kueck, Nadine A and Spacek, Petr and Cornelissen, Nicolas Vincent and Huewel, Sabine and Dieterich, Christoph and Rentmeister, Andrea},
year={2022},
month={Mar}
}