Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
J Am Chem Soc. 2012 May 16;134(19):8050-3. doi: 10.1021/ja302197x. Epub 2012 May 7.
A nuclease-resistant RNA enzyme, constructed entirely from L-ribonucleotides, was shown to undergo ligand-dependent, self-sustained replication with exponential growth. The catalytic motif is based on a previously described RNA ligase that can undergo either self- or cross-replication but had been limited in its application to ligand sensing due to its susceptibility to degradation by ribonucleases. The self-replicating RNA enzyme and its RNA substrates were prepared synthetically from either D- or L-nucleoside phosphoramidites. The D and L reaction systems undergo isothermal, ligand-dependent exponential amplification in the same manner, but only the l system is impervious to ribonucleases and can operate, for example, in the presence of human serum. This system has potential for the quantitative detection of various ligands that are present within biological or environmental samples. In addition, this work provides the first demonstration of the self-sustained exponential amplification of nonbiological molecules.
一种完全由 L-核糖核苷酸构建的耐核酸酶 RNA 酶被证明能够进行配体依赖性、自我维持的复制，实现指数级增长。该催化基序基于先前描述的 RNA 连接酶，它可以进行自我或交叉复制，但由于其易被核糖核酸酶降解，限制了其在配体检测中的应用。自我复制的 RNA 酶及其 RNA 底物可以通过 D-或 L-核苷膦酰胺合成制备。D 和 L 反应系统以相同的方式进行等温、配体依赖性指数扩增，但只有 L 系统不受核糖核酸酶的影响，可以在例如人血清存在的情况下进行反应。该系统具有定量检测生物或环境样品中存在的各种配体的潜力。此外，这项工作首次证明了非生物分子的自我维持指数级扩增。
