Sequence-specific inhibition of a transcription factor by circular dumbbell DNA oligonucleotides

The inhibition of specific transcription regulatory proteins is a new approach to control gene expression. The transcriptional activities of DNA-binding proteins can be inhibited by the use of double-stranded oligonucleotides that compete for the binding to their specific target sequences in promote...

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Veröffentlicht in:FEBS letters 1999-11, Vol.461 (3), p.136-140
Hauptverfasser: Hosoya, Takeshi, Takeuchi, Hiroaki, Kanesaka, Yoshiyuki, Yamakawa, Hidefumi, Miyano-Kurosaki, Naoko, Takai, Kazuyuki, Yamamoto, Naoki, Takaku, Hiroshi
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container_end_page 140
container_issue 3
container_start_page 136
container_title FEBS letters
container_volume 461
creator Hosoya, Takeshi
Takeuchi, Hiroaki
Kanesaka, Yoshiyuki
Yamakawa, Hidefumi
Miyano-Kurosaki, Naoko
Takai, Kazuyuki
Yamamoto, Naoki
Takaku, Hiroshi
description The inhibition of specific transcription regulatory proteins is a new approach to control gene expression. The transcriptional activities of DNA-binding proteins can be inhibited by the use of double-stranded oligonucleotides that compete for the binding to their specific target sequences in promoters and enhancers. We used nicked (NDODN-κB) and circular (CDODN-κB) dumbbell DNA oligonucleotides containing a NF-κB binding site to analyze the inhibition of the NF-κB-dependent activation of the human immunodeficiency virus type-1 (HIV-1) enhancer. The dumbbell DNA oligonucleotides are stable, short segments of double-stranded DNA with closed nucleotide loops on each end, which confer resistance to exonucleases. The dumbbell and other oligonucleotides (decoys) with the NF-κB sequence were found to compete with the native strand for NF-κB binding. The circular dumbbell and double-stranded phosphorothioate oligonucleotides competed with the native strand for binding to the NF-κB binding proteins, while the nicked NF-κB dumbbell was a less effective competitor. In Jurkat T-cells, the dumbbell and other oligonucleotides were tested for their ability to block the activation of the plasmid HIV-NL4-3 Luc. The CDODN-κB strongly inhibits the specific transcriptional regulatory proteins, as compared with the NDODN-κB and the double stranded phosphodiester oligonucleotides. On the other hand, the double stranded phosphorothioate oligonucleotides could also block this activation, but the effect was non-specific. The circular (CDODN) dumbbell oligonucleotides may efficiently compete for the binding of specific transcription factors within cells, thus providing anti-HIV-1 or other therapeutic effects.
doi_str_mv 10.1016/S0014-5793(99)01450-7
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subjects AIDS/HIV
Anti-HIV
Binding Sites - drug effects
Binding, Competitive
Decoy
DNA dumbbell
Enhancer Elements, Genetic
Gene Expression Regulation, Viral - drug effects
Genes, Reporter
HIV-1 - genetics
Humans
Infant, Newborn
Inhibition of transcription factor (NF-κB)
Jurkat cell
Jurkat Cells
Luciferase assay
Luciferases - biosynthesis
Luciferases - genetics
NF-kappa B - antagonists & inhibitors
NF-kappa B - chemistry
Nucleic Acid Conformation
Oligodeoxyribonucleotides - pharmacology
Protein Binding
Recombinant Fusion Proteins - biosynthesis
Thionucleotides - pharmacology
Transfection
title Sequence-specific inhibition of a transcription factor by circular dumbbell DNA oligonucleotides
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