Large-scale tethering of RNA binding proteins discovers alternative splicing regulators [RIP-Seq]

To elucidate the role of large sets of RBPs, we developed a comprehensive and diverse plasmid resource3 of 765 RBPs fused to the MS2 bacteriophage coat protein. When systematically tethered to an intronic 4 MS2 hairpin downstream ofan alternative exon, we identified 55 RBPs that significantly induced exon 5 retention. Overall design: RBPs were exogeneously expressed and immunoprecipitated by affinity tag and NGS was performed.