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. 1993 Jun 25;21(12):2789–2796. doi: 10.1093/nar/21.12.2789

In vivo transcription of a progesterone-responsive gene is specifically inhibited by a triplex-forming oligonucleotide.

N H Ing 1, J M Beekman 1, D J Kessler 1, M Murphy 1, K Jayaraman 1, J G Zendegui 1, M E Hogan 1, B W O'Malley 1, M J Tsai 1
PMCID: PMC309654  PMID: 8332487

Abstract

Oligonucleotides provide novel reagents for inhibition of gene expression because of their high affinity binding to specific nucleotide sequences. We describe a 38 base, single-stranded DNA that forms a triple helix or 'triplex' on progesterone response elements of a target gene. This triplex-forming oligonucleotide binds with a Kd = 100 nM at 37 degrees C and physiological pH, and blocks binding of progesterone receptors to the target. Furthermore, it completely inhibited progesterone receptor-dependent transcription in vitro. To approach in vivo conditions, triplex-forming oligonucleotides were tested in cell transfection studies. The derivation of the oligonucleotides with cholesterol enhanced their cellular uptake and nuclear concentration by at least four-fold. The cholesterol-derivatized triplex-forming oligonucleotide specifically inhibited transcription of the PRE-containing reporter gene in cells when applied to the medium at micromolar concentrations. This is the first demonstration of steroid-responsive gene inhibition by triplex formation and joins the growing body of evidence indicating that oligonucleotides have therapeutic potential.

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Selected References

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