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.
Department of Pediatrics, Childrens Hospital, Los Angeles, CA 90054.
H
H
Wu
J S
JS
Holcenberg
J
J
Tomich
J
J
Chen
P A
PA
Jones
S H
SH
Huang
K L
KL
Calame
D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D004742:Enhancer Elements, Genetic; D015870:Gene Expression / Q000187:drug effects; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000037:antagonists & inhibitors; D015183:Restriction Mapping; D014158:Transcription, Genetic / Q000187:drug effects*
D009838:Oligodeoxyribonucleotides; D016372:RNA, Antisense; D012333:RNA, Messenger; D012313:RNA