In vivotranscription of a progesterone-responsive gene is specifically inhibited by a triplex-forming oligonucleotide

作者：Nancy H. Ing, Johanna M. Beekman, Donald J. Kessler, Mark Murphy, Krishna Jayaraman, Joseph G. Zendegui, Michael E. Hogan, Bert W. O’Malley, Ming-Jer Tsai · 发表于：Nucleic Acids Research · 年份：1993 · DOI：10.1093/nar/21.12.2789 · 被引用次数：92 · 研究领域：DNA and Nucleic Acid Chemistry、RNA Interference and Gene Delivery、Estrogen and related hormone effects

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.