Targeting cytoplasmic cap methylation

To identify targets of cytoplasmic capping, we developed an inducible U2OS cell line overexpressing a dominant negative cytoplasmic form of cap methyltransferase. Overexpressing this dominant negative protein results in a lack of methylation on cytoplasmically recapped RNAs and thus their degradation through the cells quality control mechanisms. Comparing RNA expression in induced and uninduced cell lines thus reveals targets of cytoplasmic recapping.