Part:BBa_K1942000:Design

RNA interference (RNAi) is an emerging technology of gene silencing. It uses siRNA segments to destroy specific mRNA, thereby, shut down the expression of target genes. RNAi has been demonstrated as a novel treatment modality of cancer. In our project, we choose lung cancer as the disease model. The activation of KRAS oncogene is specifically associated with lung cancer, so we picked KRAS as our target oncogene. Then we designed anti-KRAS siRNAs as therapeutic agents to degrade KRAS mRNA, therefore, silencing K-ras protein expression and function. We used a professional software designed by SYSU-software to find the ideal siRNA binding sequence to insure the specificity and efficiency of KRAS knock-down. This tool was also used to design pairs of oligonucleotides needed to generate short hairpin RNAs (shRNAs) from the plasmid. When the shRNA plasmids of KRAS are transfected into HEK293 cells, the dsRNA is cleaved into siRNA by the enzyme Dicer, and then target the mRNA of KRAS, thus effectively silencing the KRAS gene.

Figure 1. The sequence of KRAS shRNA