Novel kethoxal derivatives-assisted identification of single-stranded RNA-binding proteins

This project introduces the kethoxal-assisted single-stranded RNA interactome capture (KASRIC) strategy to enable transcriptome-wide identification of single-stranded RBPs (ssRBPs). The kethoxal probe selectively modifies unpaired guanine residues in RNA single-stranded regions, thereby blocking ssRBP interaction near these sites. After RNA-protein crosslinking and oligo(dT) bead-mediated capture, enriched proteins are analyzed using DIA proteomics. Proteins with significantly down-regulated abundance in kethoxal-treated groups (N-Block and A-Block) compared to the crosslinked control group (cCL) are identified as candidate ssRBPs.