CSDE1 promotes passenger strand cleavage of miR-486

Strand separation of the RNA duplex is a critical step in miRNA biogenesis, in which only the guide strand is retained to form the mature miRISC. The erythroid miRNA miR-486-5p uniquely requires AGO2 catalytic slicing of its passenger strand miR-486-3p, yet the factors regulating this process remain poorly understood. Here, we identify the RNA-binding protein CSDE1 as a key cofactor that promotes AGO2-dependent passenger-strand removal and miR-486-5p maturation. CSDE1 interacts with the N-terminal domain of AGO2, essential for strand release. Loss of CSDE1 increased the miR-486-3p/5p ratio, reduced cleavage efficiency, and derepressed miR-486 targets such as FOXO1 and PTEN. Restoring full-length CSDE1 expression restored AGO2-dependent passenger strand cleavage and this function requires the N-terminal cold-shock domain CSD1, which mediates CSDE1 interaction with AGO2. Together, these results demonstrate CSDE1 as a critical regulator of AGO2-mediated strand separation, potentiating miR-486-5p function in target gene silencing associated with in hematological malignancies.