Ainara Castellanos, Ikerbasque researcher at the UPV/EHU, has led a study that has been published in the prestigious journal Scientific Reports in which a new RT-QPCR approach for the relative quantification of the specific residue of m6A methylation of the RNA of immunological genes is described.
N6-methyladenosine (m6A) is the most common and abundant chemical modification in RNA molecules. It has been recently described that these modifications are involved in the regulation of different biological processes including the immune response. Different methods have been developed in order to map and quantify m6A marks, but the quantification of m6A levels in certain locations of the RNA has been technically challenging and the existing protocols are long and tedious and often involve next-generation sequencing.
In this paper, it is described a novel and sinple RT-QPCR based approach for the relative quantification of candidate m6A regions that takes advantage of the diminished capacity of BstI enzyme to retrotranscribe m6A residues. Using this technique, the recently described m6A methylation in the 3′UTR of SOCS1 and SOCS3 transcripts have been confirmed. Moreover, using the method presented here, alterations in the relative levels of m6A in specific motifs of SOCS genes in celiac disease patients and in pancreatic β-cells exposed to inflammatory stimuli have been observed.
For more information: https://rdcu.be/bqC8Q