Department of Regulatory Bioorganic Chemistry, The Institute of Scientific and Industrial Research, Osaka University, Ibaraki 567-0047, Osaka, Japan.
J Am Chem Soc. 2010 Aug 18;132(32):11171-8. doi: 10.1021/ja1032088.
Guanine quadruplex structures in DNA and RNA affect normal cellular processes such as replication, recombination, and translation. Thus, controlling guanine quadruplex structures could make it possible to manipulate the biological function of nucleic acids. Here, we report a novel antisense strategy using guanine-tethered antisense oligonucleotides (g-ASs) that introduces an RNA-DNA heteroquadruplex structure on RNA templates in a predictable and sequence-specific manner, which in practice effectively inhibited reverse transcription on a variety of RNA sequences, including the HIV-1 RNA genome. Reverse transcriptase-mediated enzymatic analysis, together with other biophysical analyses, elucidated a cooperative binding of duplex and quadruplex in g-AS-RNA complexes. The remarkable ability of g-ASs to inhibit reverse transcription could make possible the development of novel anti-retroviral gene therapies based on blocking the replication of RNA genomes to complementary DNA, which is a critical step for integration into the host's genome.
DNA 和 RNA 中的鸟嘌呤四链体结构会影响正常的细胞过程，如复制、重组和翻译。因此，控制鸟嘌呤四链体结构可以实现对核酸生物功能的操控。在这里，我们报告了一种新的反义策略，使用鸟嘌呤连接的反义寡核苷酸（g-ASs），以可预测且序列特异性的方式在 RNA 模板上引入 RNA-DNA 杂四链体结构，实际上有效地抑制了多种 RNA 序列的逆转录，包括 HIV-1 RNA 基因组。逆转录酶介导的酶分析以及其他生物物理分析阐明了双链体和四链体在 g-AS-RNA 复合物中的协同结合。g-ASs 抑制逆转录的显著能力可能使基于阻断 RNA 基因组复制互补 DNA 的新型抗逆转录病毒基因疗法的开发成为可能，这是整合到宿主基因组的关键步骤。
