Role of mRNA capping in ESCs and early embryo development (360G-Wellcome-109111_Z_15_A)

Pluripotency maintenance and transition to differentiation require tight regulation of gene expression at the transcriptional and post-transcriptional level. The mRNA cap consists of a series of co-transcriptional modifications of the nascent transcript that are important for its processing and translation initiation. The mature cap structure requires methylation by the complex RNMT-RAM. A recent study showed that RAM, the mRNA cap methyltransferase activating subunit, is highly expressed in embryonic stem cells (ESCs) and required for the expression of pluripotency-associated genes such as Oct4 and Sox2. During the process of neurodifferentiation, degradation of RAM is required for the loss of pluripotency and expression of neural-associated genes. Given the importance of mRNA cap methylation in pluripotency maintenance, in this project we will investigate the role of mRNA capping in the process of ESCs differentiation and early embryo development. We will analyse the expression patterns of the capping enzymes in both embryoid bodies (EBs) and blastocysts along with pluripotency and lineage-specific markers. We will perform this analysis in wild-type and capping enzyme knockout-derived EBs and blastocysts. Depending on the observed expression patterns, we will pursue any interesting emerging phenotypes that may suggest the involvement of regulated mRNA capping in the differentiation process.