Construction of Eukaryotic Expression Vector of Short Hairpin RNA for ezrin

Objective To construct the short hairpin RNA （shRNA） expression vector specific for ezrin. Methods Oligonucleotides were designed specific for ezrin gene. After annealing, the formed double-stranded DNAs were ligated with pSUPER. The expression vectors of pSUPER -shRNA were identified by enzyme digestion and sequence analysis and transfected into MCF-7 cells. The expression of ezrin was analyzed by RT-PCR and Western blot. Results The vector was identified by restriction enzyme digestion and sequence analysis. The expression vectors of pSUPER-shRNA was successfully constructed and transfected into MCF-7 cells. The ezrin expression was significantly suppressed after transfection with ezrin shRNA vector. Conclusion The pSUPER -shRNA specific for ezrin is successfully constructed and it will be helpful for further study on the significance of ezrin on the metastasis of breast cancer.