RNA Methylation Activity/Inhibition

The formation of m⁶A RNA appears to be a co-transcriptional event taking place early on in the RNA lifecycle and is mediated by a multi-protein methyltransferase complex composed, in part, of METTL enzymes. Chemical addition of a methyl group at the nitrogen-6 position of adenosine (A) residues to form m⁶A occurs via the METTL3/14 heterodimer. Through FTO-mediated oxidative demethylation, m⁶A is converted in a step-wise manner to hm⁶A and subsequently f6A before finally reverting back to A. The recent discoveries of these m⁶A methylase “writers” and their associated demethylase “erasers” in mammals uncovered the reversibility of the m⁶A modification, exposing potential therapeutic targets for m⁶A dysregulation-related diseases.

EpigenTek’s ELISA-based Epigenase assays provide rapid measures of m⁶A methylase and demethylase activity levels from cell/tissue nuclear extracts or purified enzymes in a high-throughput format. A unique m⁶A substrate is stably coated on the assay wells. Bioactive enzymes from input samples will transfer methyl groups to (methylases), or remove them from (demethylases), the bound substrate. Substrate m⁶A methylation is subsequently detected by a high-affinity antibody specific for this modification. Quantitative measurements of methylase/demethylase activity and inhibition (for enzyme inhibitor screening) can be quickly obtained within only a few hours.