REGULATION OF GENE-EXPRESSION WITH DOUBLE-STRANDED PHOSPHOROTHIOATE OLIGONUCLEOTIDES
REGULATION OF GENE-EXPRESSION WITH DOUBLE-STRANDED PHOSPHOROTHIOATE OLIGONUCLEOTIDES
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DOI：
10.1126/science.2237444
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发表时间：
1990-11-16
期刊：
影响因子：
56.9
通讯作者：
NABEL, GJ
中科院分区：
文献类型：
--
作者：
BIELINSKA, A;SHIVDASANI, RA;NABEL, GJ
通过抑制特定转录调节蛋白来改变基因转录对于确定这些因子如何参与细胞分化是必要的。这些蛋白质的功能可以通过多种方法来拮抗，每种方法都有特定的局限性。使用含有八聚体或κB共有序列的双链(ds)硫代磷酸寡核苷酸实现了对序列特异性DNA结合蛋白的抑制。硫代磷酸酯寡核苷酸特异性结合八聚体转录因子或核因子(NF)-κB。修饰的寡核苷酸比标准双链寡核苷酸更有效地在细胞中积累，并以特定方式调节基因表达。当将适当的硫代磷酸酯寡核苷酸添加至瞬时转染的B细胞系时，报告质粒的八聚体依赖性激活或人类免疫缺陷病毒(HIV)增强子的NF-κB依赖性激活被抑制。向 Jurkat T 白血病细胞中添加含有八聚体共有序列的硫代磷酸寡核苷酸可抑制白细胞介素 2 (IL-2) 分泌，其程度与在 IL-2 增强子中观察到的突变八聚体位点相似。双链硫代磷酸寡核苷酸可能竞争结合特定转录因子，并可提供抗病毒、免疫抑制或其他治疗作用。
Alteration of gene transcription by inhibition of specific transcriptional regulatory proteins is necessary for determining how these factors participate in cellular differentiation. The functions of these proteins can be antagonized by several methods, each with specific limitations. Inhibition of sequence-specific DNA-binding proteins was achieved with double-stranded (ds) phosphorothioate oligonucleotides that contained octamer or .kappa.B consensus sequences. The phosphorothioate oligonucleotides specifically bound either octamer transcription factor or nuclear factor (NF)-.kappa.B. The modified oligonucleotides accumulated in cells more effectively than standard ds oligonucleotides and modulated gene expression in a specific manner. Octamer-dependent activation of a reporter plasmid or NF-.kappa.B-dependent activation of the human immunodeficiency virus (HIV) enhancer was inhibited when the appropriate phosphorothioate oligonucleotide was added to a transiently transfected B cell line. Addition of phosphorothioate oligonucleotides that contained the octamer consensus to Jurkat T leukemia cells inhibited interleukin-2 (IL-2) secretion to a degree similar to that observed with a mutated octamer site in the IL-2 enhancer. The ds phosphorothioate oligonucleotides probably compete for binding of specific transcription factors and may provide anti-viral, immunosuppressive, or other therapeutic effects.