Clusel C, Meguenni S, Elias I, Vasseur M, Blumenfeld M
Genset, Paris, France.
Gene Expr. 1995;4(6):301-9.
Transcriptional control in eukaryotes results from the interplay between DNA sequences in promoters, enhancers, or silencers and transcription factors. Selective control of gene expression can thus be achieved by inhibiting specific transcription factor/DNA interactions. Transcriptional activity of DNA binding transcription factors can be inhibited by competition with double-stranded oligonucleotides (decoys) that contain their specific recognition sequences. The immediate early protein ICP4 of herpes simplex virus type 1 (HSV-1) is a sequence-specific DNA binding protein that is essential for viral replication. We synthesized double-stranded hairpin phosphodiester oligonucleotides carrying ICP4 sites and demonstrated their ability to specifically titrate ICP4. Upon addition to Vero cells, ICP4 hairpin decoys significantly reduced HSV-1 titers (IC50 = 0.3 microM), whereas a control hairpin oligonucleotide had no activity. Antiviral activity of ICP4 hairpin decoys was correlated to their relative binding affinities. These results show that phosphodiester oligonucleotides can compete for binding of specific transcription factors within cells, thus providing a potential therapeutic tool to control disease-causing genes.
真核生物中的转录调控源于启动子、增强子或沉默子中的DNA序列与转录因子之间的相互作用。因此，通过抑制特定转录因子/DNA相互作用，可以实现对基因表达的选择性控制。与包含其特定识别序列的双链寡核苷酸（诱饵）竞争，可抑制DNA结合转录因子的转录活性。单纯疱疹病毒1型（HSV-1）的立即早期蛋白ICP4是一种序列特异性DNA结合蛋白，对病毒复制至关重要。我们合成了携带ICP4位点的双链发夹磷酸二酯寡核苷酸，并证明了它们特异性滴定ICP4的能力。将其添加到Vero细胞中后，ICP4发夹诱饵显著降低了HSV-1滴度（IC50 = 0.3 microM），而对照发夹寡核苷酸则无活性。ICP4发夹诱饵的抗病毒活性与其相对结合亲和力相关。这些结果表明，磷酸二酯寡核苷酸可以在细胞内竞争特定转录因子的结合，从而提供了一种控制致病基因的潜在治疗工具。
