N6-methyladenosine (m6A) profiling of EndoC-bH1 cell line and RNA seq of Mettl14 knockout mice beta cell

In type 2 diabetes, pancreatic beta-cells fail to compensate for the presence of insulin resistance in target tissues and represent a central player in the disease development. Identifying and studying innovative molecular mechanisms that lead to beta-cell failure in diabetes represent an interesting line of research and are necessary. N6-Methyladenosine (m6A) is the most abundant modification in mRNA and is found virtually in all mammals. Through m6A-profiling of m6A methyltransferase depleted animal model and human beta cell model, we aim to characterize the pathways affected by m6A methylation in the beta cells. Total RNA was extracted from the tissue by TRIzol. mRNA was enriched by polyT beads followed by fragmentation with Bioruptor ultrasonicator. m6A-immunoprecipitation were performed using EpiMark N6-Methyladenosine enrichment kit (NEB cat. E1610S). Kapa RNA hyper kit for Illumina was used to construct library from mRNA for input RNA and eluted RNA from the m6A-IP. The libraries were sequenced by the Hiseq4000 platform at SE50 mode.