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Although numerous approaches have been developed to map RNA-binding sites of individual RNA-binding proteins (RBPs), few methods exist that allow assessment of global RBP-RNA interactions. Here, we describe PIP-seq, a universal, high-throughput, ribonuclease-mediated protein footprint sequencing approach that reveals RNA-protein interaction sites throughout a transcriptome of interest. We apply PIP-seq to the HeLa transcriptome and compare binding sites found using different cross-linkers and ribonucleases. From this analysis, we identify numerous putative RBP-binding motifs, reveal novel insights into co-binding by RBPs, and uncover a significant enrichment for disease-associated polymorphisms within RBP interaction sites. © 2014 Silverman et al.; licensee BioMed Central Ltd.
Silverman, I. M., Li, F., Alexander, A., Goff, L., Trapnell, C., Rinn, J. L., & Gregory, B. D. (2014). RNase-mediated protein footprint sequencing reveals protein-binding sites throughout the human transcriptome. Genome Biology, 15(1). https://doi.org/10.1186/gb-2014-15-1-r3