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Construction of RNAi Adenovirus Targeting both p53 and VEGF Synchronously and Its Gene Silencing Function
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Abstract:
Objective : To construct RNAi adenovirus targeting both VEGF and p53 in the same time. Methods: The interfering plasmids targeting VEGF and p53 were constructed respectively. Then the H1 promoter and RNAi sequence were cut and ligated to pAdTrack successively to generate pAdTrack/VEGF/p53, which was transfected into BJ5183 cells containing pAdEasy-1 to generate pAd/VEGF/p53 after homologous recombination, harvesting the adenovirus after infecting 293 cells. The knockdown of VEGF and p53 by pAd/VEGF/p53 was detected through real-time PCR after infecting MCF-7 cells. Results:The constructed pAd/VEGF/p53 containing being inserted two H1 promoters and RNAi sequences was identified by BamH digesting. The VEGF and p53 mRNA level of MCF-7 cells decreased obviously after being infected by pAd/VEGF/p53. Conclusions: The RNAi adenovirus targeting both VEGF and p53 in the same time was successfully constructed and it can silence both genes obviously.
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