Fluorogenic and Bioorthogonal Modification of RNA Using Photoclick Chemistry
Fluorogenic and Bioorthogonal Modification of RNA Using Photoclick Chemistry
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DOI：
10.3390/biom10030480
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发表时间：
2020-03-01
期刊：
影响因子：
5.5
通讯作者：
Wagenknecht, Hans-Achim
中科院分区：
文献类型：
--
作者：
Krell, Katja;Wagenknecht, Hans-Achim
合成了一种溴芳基四唑修饰的尿苷，作为一种新的RNA构建块，可用于生物正交、光活化和合成后的荧光染料修饰。用马来酰亚胺共轭荧光团在预合成RNA的内部和末端位置进行光（300 nm LED）照射，可实现“光点击”型修饰，产量高。用三种不同的染料证明了该反应。在辐照过程中，由于固有荧光吡唑啉部分作为光点击产物的形成，发射增加。反应产物吡唑啉（弱发射体）与光点击染料（强发射体）之间的能量传递显著增强了光点击反应的荧光性。RNA-染料缀合物具有显著的荧光特性，特别是荧光强度可提高9.4倍，这对细胞内RNA的化学生物学和荧光成像具有重要意义。
A bromoaryltetrazole-modified uridine was synthesized as a new RNA building block for bioorthogonal, light-activated and postsynthetic modification with commercially available fluorescent dyes. It allows "photoclick"-type modifications by irradiation with light (300 nm LED) at internal and terminal positions of presynthesized RNA with maleimide-conjugated fluorophores in good yields. The reaction was evidenced for three different dyes. During irradiation, the emission increases due to the formation of an intrinsically fluorescent pyrazoline moiety as photoclick product. The fluorogenecity of the photoclick reaction was significantly enhanced by energy transfer between the pyrazoline as the reaction product (poor emitter) and the photoclicked dye as the strong emitter. The RNA-dye conjugates show remarkable fluorescent properties, in particular an up to 9.4 fold increase of fluorescence, which are important for chemical biology and fluorescent imaging of RNA in cells.