Fig. 1

Argonaute crosslinking and immunoprecipitation experiments enable the high-throughput capturing of miRNA targets. a Illustration of miRNA targeting. miRNAs are loaded on AGO2 and guide the RISC complex to target MRE(s). RISC binding to its target genes can either cease their translation or induce their cleavage and/or degradation. b Peaks derived from 5 AGO-PAR-CLIP libraries on HEK293 cells and from 3 non-RBP background libraries are presented for T-to-C and non-T-to-C AGO-bound regions. The red-and-blue vertical lines represent T-to-C transition sites. Both types of AGO-enriched clusters are clearly distinguished from background signal. Chimeric miRNA-target fragments overlap with (non-)T-to-C peaks providing direct validation for specific miRNA-target pairs (hsa-miR-19a-3p–Ran and hsa-miR-103a-3p–Rps14). microCLIP identifies the aforementioned interactions as a 7-mer (chr12:131,361,200–131,361,400, Ran gene 3′ UTR) and an 8-mer with a 3′ compensatory site (chr5:149,826,350–149,826,550, Rps14 gene CDS), respectively. The 3D depictions of AGO2 were based in the PDB structure 5JS1