Ganz Dorothée, Harijan Dennis, Wagenknecht Hans-Achim
Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT) Fritz-Haber-Weg 6 76131 Karlsruhe Germany
RSC Chem Biol. 2020 Jun 2;1(3):86-97. doi: 10.1039/d0cb00047g. eCollection 2020 Aug 1.
Labelling of nucleic acids as biologically important cellular components is a crucial prerequisite for the visualization and understanding of biological processes. Efficient bioorthogonal chemistry and in particular cycloadditions fullfill the requirements for cellular applications. The broadly applied Cu(i)-catalyzed azide-alkyne cycloaddition (CuAAC), however, is limited to labellings and in fixed cells due to the cytotoxicity of copper salts. Currently, there are three types of copper-free cycloadditions used for nucleic acid labelling in the cellular environment: (i) the ring-strain promoted azide-alkyne cycloaddition (SPAAC), (ii) the "photoclick" 1,3-dipolar cycloadditions, and (iii) the Diels-Alder reactions with inverse electron demand (iEDDA). We review only those building blocks for chemical synthesis on solid phase of DNA and RNA and for enzymatic DNA and RNA preparation, which were applied for labelling of DNA and RNA or , in the cellular environment, in fixed or in living cells, by the use of bioorthogonal cycloaddition chemistry. Additionally, we review the current status of orthogonal dual and triple labelling of DNA and RNA to demonstrate their potential for future applications or .
将核酸标记为重要的生物细胞成分是可视化和理解生物过程的关键前提。高效的生物正交化学，特别是环加成反应，满足了细胞应用的要求。然而，由于铜盐的细胞毒性，广泛应用的铜（I）催化的叠氮化物-炔烃环加成反应（CuAAC）仅限于标记固定细胞。目前，在细胞环境中用于核酸标记的无铜环加成反应有三种类型：（i）环张力促进的叠氮化物-炔烃环加成反应（SPAAC），（ii）“光点击”1,3-偶极环加成反应，以及（iii）具有反向电子需求的狄尔斯-阿尔德反应（iEDDA）。我们仅综述那些用于DNA和RNA固相化学合成以及酶促DNA和RNA制备的构建模块，这些构建模块通过生物正交环加成化学用于在细胞环境中、固定或活细胞中标记DNA和RNA。此外，我们综述了DNA和RNA正交双标记和三标记的现状，以展示它们未来应用的潜力。
