ADAMTSL3 - a potential direct target of miR-144-3p in chondrocyte cell

Abstract

The miR-144-3p was discovered to upregulate in osteoarthritis. However, the function of this molecule is still poorly understood. Identification of microRNA target genes could shed a light on miRNA functions. In this project, we aim to identify a direct target of miR-144-3p. Potential targets of miR-144-3p were predicted by bioinformatic algorithms. Expression of ADAMTSL3 in miRNA-144-3p gain-and- loss- of function experiment was quantified by Realtime-PCR. Next, the 3’UTR of ADAMTSL3 containing miR-144-3p-binding sites was subcloned in the 3’;UTR of the Firefly luciferase gene on the pmiRGlo. To test the complementary binding of miR-144-3p to ADAMTSL3 mRNA, we employed the luciferase assay using pmiRGlo-ADAMTSL3 vector or pmiRGlo-ADAMTSL3 mutant in which miR-144-3p binding sites in the 3’UTR was mutated. Results showed that miRNA-144-3p directly inhibits the expression of ADAMTSL3 through its ability to pair at “seed” sites. This study provides evidence for the interaction of miRNA-144-3p and its target, ADAMTSL3 mRNA, in chondrocytes.