Construction and validation of adenovirus expressing dominant negative TGF-β type Ⅰ receptors
AIM: To construct recombinant adenovirus containing dominant negative TGF-β type Ⅰ receptors and to validate its inhibitory effect on the ligand's function. METHODS: The adenovirus containing dominant negative TGF-β type Ⅰ receptors is constructed using molecular clone and recombinant adenovirus assay. C3H10 cells were then infected by the adenovirus and the expression of adenovirus in C3H10 cells was confirmed by fluorescence microscope and RT-PCR. Alkaline phosphatase quantitative assay and luciferase reporter assay were used to analyze the inhibitory effect of the adenovirus on BMP2-induced ALP activity and TGF-β1-induced luciferase activity. RESULTS: The adenovirus containing dominant negative TGF-β type Ⅰ receptors infected C3H10 cells and expressed dominant negative TGF-β type Ⅰ receptors in C3H10 cells. Adenovirus expressing dominant negative TGF-β type Ⅰ receptors inhibited the BMP2-induced ALP activity and TGF-β1-induced luciferase activity. CONCLUSION: The adenovirus containing dominant negative TGF-β type Ⅰ receptors is successfully constructed and its inhibitive effect on the ligand's function is confirmed, which provides a useful approach to further studying the functions of TGF-β type Ⅰ receptors and to targeting TGF-β signaling for the disease therapy.
NETL
NSTL
万方数据
