Shinohara Ken-ichi, Sasaki Shunta, Bando Toshikazu, Sugiyama Hiroshi
Graduate School of Science, Kyoto University, Kitashirakawa Oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
Nucleic Acids Symp Ser (Oxf). 2005(49):75-6. doi: 10.1093/nass/49.1.75.
We have demonstrated that hairpin pyrrole (Py)-imidazole (Im) polyamide-CPI conjugates selectively induced luciferase gene silencing by sequence-specific alkylation of the coding region. Recently, we developed a new type of Py-Im polyamide CBI conjugate with an indole linker as a stable sequence-specific alkylating agent. In this study, we investigated the gene silencing ability of polyamides A, B and C, which potentially target specific sequences in the promoter region, non-coding strand, and coding strand of the green fluorescent protein (GFP) gene, respectively. The GFP vectors were transfected into human colon carcinoma cells (HCT116), and the cells treated with 100 nM of the polyamides for 24 h. Using direct observation of cell by fluorescence microscopy, a significant GFP-gene silencing effect was only seen with treatment with polyamide C. Polyamides A and B did not show such activity.
我们已经证明，发夹状吡咯（Py）-咪唑（Im）聚酰胺-CPI缀合物通过对编码区进行序列特异性烷基化，选择性地诱导荧光素酶基因沉默。最近，我们开发了一种新型的带有吲哚连接体的Py-Im聚酰胺CBI缀合物，作为一种稳定的序列特异性烷基化剂。在本研究中，我们研究了聚酰胺A、B和C的基因沉默能力，它们分别可能靶向绿色荧光蛋白（GFP）基因启动子区、非编码链和编码链中的特定序列。将GFP载体转染到人结肠癌细胞（HCT116）中，并用100 nM的聚酰胺处理细胞24小时。通过荧光显微镜直接观察细胞，仅在用聚酰胺C处理时观察到显著的GFP基因沉默效应。聚酰胺A和B没有显示出这种活性。
