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通过核酸模板反应进行扩增。

Amplification by nucleic acid-templated reactions.

作者信息

Michaelis Julia, Roloff Alexander, Seitz Oliver

机构信息

Institut für Chemie der Humboldt-Universität zu Berlin, Brook-Taylor-Strasse 2, 12489-Berlin, Germany.

出版信息

Org Biomol Chem. 2014 May 14;12(18):2821-33. doi: 10.1039/c4ob00096j.

Abstract

Nucleic acid-templated reactions enable the design of conditional reaction systems, in which bond formation occurs only when a particular DNA or RNA molecule is present. Such reaction systems are currently being explored for applications in DNA/RNA diagnosis, drug screening and as a means to design gene expression specific therapy. However, biological nucleic acid templates usually have low abundance. Therefore, either the targeted nucleic acid template has to be multiplied by means of an amplification step or the template itself has to act as a catalyst which amplifies product formation. This critical review highlights the recent advancements in nucleic acid-templated reactions that proceed with turnover in template and thereby provide a means of amplification. Improvements in reaction engineering and the development of new chemistries have pushed the limits from 10(1) to 10(2)-10(3) turnovers. This includes reaction systems that lead to the ligation of oligonucleotides or to the interconversion of appended functional groups beyond ligation as well as templated chemistries that enable the activation of catalysts for subsequent triggering of reactions between non-nucleotidic substrates. The present limitations and future opportunities are discussed.

摘要

核酸模板反应能够实现条件反应系统的设计,在该系统中,只有当特定的DNA或RNA分子存在时才会发生键的形成。目前正在探索此类反应系统在DNA/RNA诊断、药物筛选中的应用,以及作为设计基因表达特异性疗法的一种手段。然而,生物核酸模板的丰度通常较低。因此,要么必须通过扩增步骤来增加目标核酸模板的数量,要么模板本身必须充当催化产物形成扩增的催化剂。这篇综述重点介绍了核酸模板反应的最新进展,这些反应在模板周转的情况下进行,从而提供了一种扩增手段。反应工程的改进和新化学方法的发展已将反应极限从10¹提高到10² - 10³次周转。这包括导致寡核苷酸连接或连接以外附加官能团相互转化的反应系统,以及能够激活催化剂以随后引发非核苷酸底物之间反应的模板化学方法。本文还讨论了当前的局限性和未来的机遇。

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