Verification of smad3 targets miRNA by luciferase assay

SMAD3 gene encodes a protein that belongs to the SMAD family. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein functions as a transcriptional modulator activated by transforming growth factor-beta (TGF-beta) and is thought to play a role in the regulation of carcinogenesis. Mutations in SMAD3 gene have been associated with patients of type I Loeys-Dietz syndrome, idiopathic pulmonary fibrosis,aneurysms-osteoarthritis syndrome (AOS) and colorectal cancer (CRC). Our research shows that several different MRE formultiple miRNA is located in this region . A newly discovered Trkc-miR2 is one of them. In this study, the 3-UTR region ofSMAD3 gene and genomic fragment containing Trkc-miR2 were cloned in Psicheck-2 and PEGFP-C1,respectively. Dual luciferase test to examine the direct interaction of the miRNA and the 3-UTR was performed. The results indicate that the SMAD3 gene is a direct target of Trkc-miR2.

Keywords:

Smad3 , UTR , miRNA , Dual luciferase assay