Abstract:AIM: To observe the suppressing effect of specific small hairpin RNA(shRNA)on TGFβ-R2 expression in human lens epithelial cells(LECs).
METHODS: Specific shRNA expression vector was constructed according to the design principles of TGFβ-R2 mRNA human GeneBank siRNA synthesis and transfected into cultured 293T with TGFβ-R2 over expression vector. After effective RNAi vector was selected by Western blot, then amplification, purification, and titration. lentivirus-TGF beta-R2 shRNA was transfected into LECs, then TGFβ-R2 mRNA expression in these cells was detected by the real-time fluorescence quantitative PCR detecting system(qPCR).
RESULTS: TGFβ-R2/GV115RNAi#1 target gene expression knockdown effect is the most obvious; the titer of packaging lentivirus is 8E+8 TU/mL; the knockdown effect is significant, the interfering efficiency is 78.1%(P<0.05).
CONCLUSION: TGFβ-R2 RNAi vector was successfully constructed, and can inhibit the expression of TGFβ-R2 mRNA in human LECs.
