Inhibition of Gene Expression from the Human c-erb B Gene Promoter by a Retroviral Vector Expressing Anti-gene RNA

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Anti-gene is a potent inhibitor of transcriptional promoter activity and subsequent gene expression. Since the number of copies of DNA is much fewer than those of transcribed mRNA,anti-gene oligonucleotides targeted against the promoter region would have an advantage over antisense oligonucleotides with an mRNA as a target. This property has been exploited to suppress the expression of a variety of oncogenes for regulating tumor proliferation or viral activities. We developed a novel retroviral vector designed to express human c-erb Banti-gene RNA and to reduce the promoter activity in the cells. Mouse fibroblast NIH3T3 cells were stably transfected with an expression construct containing a truncated human c-erb B gene promoter fused to the firefly luciferase reporter gene. Addition to these cells of the c-erb B anti-gene retroviral vector targeted to the 26 bp pyrimidine-rich element in the human c-erb B gene promoter resulted in a dose-dependent decrease in the luciferase activity of the cells. The results suggest that the reduction of the luciferase activity is due to the effect of the transcribed human c-erb B anti-gene RNA on initiation of transcription of the luciferase gene. The anti-gene RNA produced by a retroviral vector is expected to suppress the expression of an appropriate oncogene and to retard the growth of tumor cells in vivo.