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. Overall design: 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.