Identification of Meiotic-P26-bound RNAs in cutlured Drosophila SL2 cells by iCLIP (individual-nucleotide resolution UV crosslinking and immunoprecipitation)
The TRIM-NHL protein Meiotic P26 (Mei-P26) governs gene expression and controls stem cell fate in Drosophila. Mei-P26 represses the translation of the key regulator protein Nanos and induces miRNA-dependent silencing to regulate stem cell renewal, differentiation and proliferation. The structural and mechanistic details of mRNA target recognition and RNA binding by its C-terminal NHL domain remain elusive. To identify its cellular target RNAs, we performed iCLIP2 experiments in cultured Drosophila SL2 cells that were transfected with Flag-tagged Mei-P26 full-length, or its NHL domain. Overall design: iCLIP2 assays were performed in three biological replicates using full-length Mei-P26 (samples 1-3) or the Mei-P26 NHL domain (samples 4-6) as bait. Reactions using non-transfected cells served as controls (samples 7-9)