Higuchi Maiko, Yamayoshi Asako, Kobori Akio, Murakami Akira
Department of Biomolecular Engineering, Kyoto Institute of Technology, Matsugasaki, Kyoto 606-8585, Japan.
Nucleic Acids Symp Ser (Oxf). 2008(52):515-6. doi: 10.1093/nass/nrn261.
Nucleic acid-based drugs, such as antisense oligonucleotide, ribozyme, and small interfering RNA, are specific compounds that inhibit gene expression at the post-transcriptional level. To develop more effective nucleic acid-based drugs, we focused on photo-reactive antisense oligonucleotides. We have optimized the structure of psoralen-conjugated oligonucleotide to improve their sequence selectivity and photo-crosslinking efficiency. Previously, we reported that photo reactive oligonucleotides containing 2'-O-psoralenyl-methoxyethyl adenosine (2'-Ps-eom) showed drastic photo-reactivity with a strictly sequence specific manner in vitro. In this report, we evaluated the binding ability toward intracellular target mRNA. The 2'-Ps-eom selectively photo-cross-linked to the target mRNA extracted from cells. The 2'-Ps-eom also cross-linked to target mRNA in cells. Furthermore, 2'-Ps-eom did not cross-link to mRNA having a mismatch base. These results suggest that 2'-Ps-eom is a powerful antisense molecule to inhibit the expression of mRNA having a point mutation.
基于核酸的药物，如反义寡核苷酸、核酶和小干扰RNA，是在转录后水平抑制基因表达的特异性化合物。为了开发更有效的基于核酸的药物，我们专注于光反应性反义寡核苷酸。我们优化了补骨脂素缀合寡核苷酸的结构，以提高其序列选择性和光交联效率。此前，我们报道了含有2'-O-补骨脂素基甲氧基乙基腺苷（2'-Ps-eom）的光反应性寡核苷酸在体外以严格的序列特异性方式表现出强烈的光反应性。在本报告中，我们评估了其对细胞内靶mRNA的结合能力。2'-Ps-eom选择性地与从细胞中提取的靶mRNA进行光交联。2'-Ps-eom也能与细胞内的靶mRNA交联。此外，2'-Ps-eom不会与具有错配碱基的mRNA交联。这些结果表明，2'-Ps-eom是一种强大的反义分子，可抑制具有点突变的mRNA的表达。
