Screening for target mRNAs of let-7a
|miRNA:||let-7a-1（duplex, 3' overhang）|
|Antibodies:||Anti-EIF2C2（AGO2）（Human）mAb（Code No. RN003M）
Mouse IgG2a（Code No. M076-3）
|Kit:||RIP-Assay Kit for microRNA（Code No. RN1005）|
Both let-7a expression levels and the AGO2-binding ratio of let-7a were increased upon transfection（Total RNA: FC（Fold change）=98, AGO2-RIP-RNA: FC=6）. Detection of other let-7a family miRNAs was considered a cross-reaction with let-7a because of a common consensus seed sequence.
The data of AGO2-RIP-RNA showed that the AGO2-binding ratio of Gene X, Y and Z were significantly changed（FC ≥ 6）. The binding status between AGO2 and mRNA was changed depending on expression level of let-7a-1.
Luciferase assay of Gene-X showed the repression of translation, which reflect on its binding status.