HyperTRIBE analysis of the m6A readers in hESCs

TRIBE/HyperTRIBE has been developed as an alternative method for transcriptome-wide mapping of the RNA targets of RBPs. This method involves fusing an RBP to the catalytic domain of the Drosophila RNA editing enzyme ADAR. When these fusion proteins are expressed in target cells, they direct A-to-I editing at residues in proximity to RNA binding sites. The control plasmid, mammalian expression plasmid, containing mCherry ADAR control and p2A GFP reporter can be obtained from Addgene (Plasmid #154786). As for RBP-TRIBE plasmid, using standard restriction enzyme-based cloning to clone RBP (DDX6, FUBP3, FXR2 or L1TD1) into TRIBE plasmid in hESCs. Overall design: All RBP-ADAR cells were analyzed with 3 duplications for HyperTRIBE.