Source was cell extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.
The binding of miRNA to mRNA was assessed using cross-linking immunoprecipitation (clip). Lysates were crosslinked, AGO1 RISC complexes were immunoprecipitated then treated with proteinase. This results in short AGO1-derived peptides cross-linked to mRNA that interfere with reverse transcription of cDNA to create small deletions, which are used to infer the region of miRNA binding to the mRNA target.
Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.
Biotinylated miRNA was transfected, allowed to assemble into RISC complexes, then used to pull down associated mRNA.
Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.
