Designing and producing of RNA interference (RNAi) in order to reduce the expression of a candidate gene involved in cancer

Abstract

Background: Cancer is one of the most important factors which affect the human health. RNA interference (RNAi) based therapies have been developed and showed significant results in this regard. One of the most important problems of siRNA system is its low stability. In order to overcome to this weakness, various types of hairpin RNA or shRNA was introduced. Over expressed colorectal cancer gene (OCC-1), which has several transcripts is considered as a hallmark of colorectal cancer. This gene has vital role in the development and progression of cancer through the cell signaling pathways, which are involved in cell proliferation. In the present study, the RNAi system was utilized in order to reduce the expression of specific transcript of OCC-1 gene. Methods: The silencer structure was designed and cloned in the expression vector pRNA-H1.1 / Neo which has a promoter for RNApolIII. Plasmid was transferred into the colorectal cancer cell line, and the expression of transcript was compared to the control samples at 24 and 72 h post transfection by Real-Time PCR. Results: The expression of transcript was reduced about 25 % at 72 h post transfection following shRNA transfection, compared to the control sample. Conclusion: The designed shRNA structure successfully reduced the expression of target gene; accordingly, it can be used in future studies in order to investigate the effect of these transcripts on development and progression of colorectal cancer.