Wagner R W, Matteucci M D, Lewis J G, Gutierrez A J, Moulds C, Froehler B C
Gilead Sciences, Inc., Foster City, CA 94404.
Science. 1993 Jun 4;260(5113):1510-3. doi: 10.1126/science.7684856.
Phosphorothioate oligodeoxynucleotides containing the C-5 propyne analogs of uridine and cytidine bind RNA with high affinity and are potent antisense inhibitors of gene expression. In a cellular assay, gene-specific antisense inhibition occurred at nanomolar concentrations of oligonucleotide, was dose-dependent and exquisitely sensitive to sequence mismatches, and was correlated with the melting temperature and length of oligonucleotide. Activity was independent of RNA target site and cell type but was detectable only when the oligonucleotides were microinjected or delivered with cell-permeabilizing agents. These oligonucleotides may have important applications in therapy and in studies of gene function.
含有尿苷和胞苷的C-5丙炔类似物的硫代磷酸酯寡脱氧核苷酸能以高亲和力结合RNA，并且是基因表达的有效反义抑制剂。在细胞试验中，基因特异性反义抑制在纳摩尔浓度的寡核苷酸时发生，呈剂量依赖性，对序列错配极为敏感，并且与寡核苷酸的解链温度和长度相关。活性与RNA靶位点和细胞类型无关，但只有当寡核苷酸通过显微注射或与细胞通透剂一起递送时才能检测到。这些寡核苷酸可能在治疗和基因功能研究中有重要应用。
