RNAi-mediated gene silencing of Bcl-2 expression

A vector used to transcribe functional short interfering RNA (siRNA) was constructed, and 64 nt oligonucleotide fragment was inserted into the downstram of the H1 promoter. Plasmids containing different bcl-2 traget sequences were transfected into Hela cells, and then the expression level of Bcl-2 was detected by indirect immunofluorescence assay (IFA) and Western Blotting. Results Vector-based RNAi had advantages over antisense RNA and synthesized siRNA because it could be delivered to the target cell more efficiently, and its effect could last longer, bcl-2 expression could be inhibited by plasmid-expressed siRNA. Three different targeting sequences were selected from bcl-2 gene, and the inhibitory effect ranged from 45% to 83.5%. Conclusion Bcl-2 expression in HeLa cells can be inhibited significantly using plasmid-based RNAi.