Ligand-dependent exponential amplification of a self-replicating L-RNA enzyme.
Ligand-dependent exponential amplification of a self-replicating L-RNA enzyme.
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DOI：
10.1021/ja302197x
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发表时间：
2012-05-16
影响因子：
15
通讯作者：
Joyce GF
中科院分区：
文献类型：
--
作者：
Olea C Jr;Horning DP;Joyce GF
一种抗核酸酶的RNA酶，完全由L-核糖核苷酸构建，被证明可以进行配体依赖性的、自我维持的复制，并呈指数增长。催化基序是基于先前描述的RNA连接酶，可以进行自我复制或交叉复制，但由于其对核糖核酸酶降解的敏感性，其在配体传感中的应用受到限制。自复制RNA酶及其RNA底物由D-或L-核苷亚磷酰胺合成制备。D和L反应系统以相同的方式进行等温的、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，并且可以在例如人血清存在下操作。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扩增。
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 non-biological molecules.