Selective multi-microrna-based gene silencing in breast cancer cells

We investigated the effectiveness of a technology that allows for the efficient silencing of three proteins in the branched-chain amino acid degradation pathway (dihydrolipoamide S acetyltransferase (DLAT), branched-chain amino acid transaminase 2 (BCAT2), and dihydrolipoamide branched-chain transacylase E2 subunit (DBT)) simultaneously. Different vectors with self-amplification, multi-miRNA, and exosome motifs were constructed. Plasmids were isolated from bacteria and transfected into breast cancer cells (BCC). Real-time PCR from isolated BCC cell RNA, as well as Western blotting of proteins, were performed. The expression of DLAT, BCAT2 and DBT genes was significantly reduced after transfection of cells with a plasmid with the complete tested RNA system. The expression of all three test proteins was inhibited in cells transfected with the full-construct SF13 plasmid.