Domnick Christof, Eggert Frank, Kath-Schorr Stephanie
Life and Medical Sciences Institute, University of Bonn, Bonn, Germany.
Chem Commun (Camb). 2015 May 14;51(39):8253-6. doi: 10.1039/c5cc01765c.
Inverse electron demand Diels-Alder cycloadditions have proven to be extremely useful for mild and additive-free orthogonal labeling of biomolecules, amongst others, for RNA labeling in vitro and in a cellular context. Here we present a method for site-specific introduction of an alkene modification into RNA via T7 in vitro transcription. For this, an unnatural, hydrophobic base pairing system developed by Romesberg and coworkers was modified introducing one or two norbornene moieties at predefined positions into RNA oligonucleotides in an in vitro transcription reaction. This allows post-transcriptional functionalization of these RNA molecules with tetrazine derivatives containing for instance fluorophores or biotin.
逆电子需求狄尔斯-阿尔德环加成反应已被证明对生物分子的温和且无添加剂的正交标记极为有用，尤其是用于体外和细胞环境中的RNA标记。在此，我们展示了一种通过T7体外转录将烯烃修饰位点特异性引入RNA的方法。为此，对Romesberg及其同事开发的一种非天然疏水碱基配对系统进行了修饰，在体外转录反应中将一个或两个降冰片烯部分引入到RNA寡核苷酸的预定义位置。这使得这些RNA分子能够用例如含有荧光团或生物素的四嗪衍生物进行转录后功能化。
