Construction Of SiRNA Expressing Vector Systerm Of Inhibiting HPV16 E6/E7 Activity

Objective To construct recombinant expressing vector of siRNA in order to inhibit HPV16 E6/E7 avtivitity. Which will be useful for the study of HPV16 E6/E7 activity inhibition and offer therapeutic means for treatment of cervical cancer.Methords 60bp DNA sequences for hairpin RNA expressing which targeted HPV16 E6/E7 gene were chemically synthesized and annealed . The pSUPER.retro vector was digested with Bgl â…¡ and Hindâ…¢ . Finally, annealed oligos were inserted into the downstream of treated pSUPER.retro' s polâ…¢ H1 promoter to construct RNAi vector by DNA recombinant technique. A circular vector containing no oligo insert was used as a negative control.Results Recombinant pSUPER-E6/E7 vectors were identified by digestion with EcoR â…  and Hindâ…¢ and confirmed by sequencing analysis. The results demonstrated that 60bp had been inserted the expected site. Furthermore, the insertion sequence were exactly correct.Conclusion siRNA expression vectors targeting HPV16 E6/E7 are successfully constructed.