Inhibition of in vitro transcription by specific double-stranded oligodeoxyribonucleotides

A potential new therapeutic approach to control gene expression is the use of double-stranded (ds) oligodeoxyribonucleotides (oligos) to compete for the binding of nuclear factors to specific promoter and enhancer elements. As a model, we have tested the effect of oligo length, sequence and number of nuclear factor binding sites on in vitro transcription of adenovirus (Ad)E1b. Short ds oligos containing an SP1-binding sequence (sp1) inhibited transcription of E1b by more than 90%. Oligos containing multiple sp1 sequences were more effective inhibitors than would be expected for a comparable number of unlinked sp1 sites. A ds oligo with phosphorothioate (PS) linkages inhibited transcription at one-tenth the concentration needed for its normal homologue. An oligo with sp1 and a consensus TATA site was no more effective than one with sp1 alone. The stability of the PS-linked oligos will allow testing of this approach in vivo if they are adequately incorporated into whole cells.