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A dimerization-based fluorogenic dye-aptamer module for RNA imaging in live cells.
Nature Chemical Biology ( IF 12.9 ) Pub Date : 2019-10-21 , DOI: 10.1038/s41589-019-0381-8 Farah Bouhedda 1 , Kyong Tkhe Fam 2 , Mayeul Collot 2 , Alexis Autour 1 , Stefano Marzi 1 , Andrey Klymchenko 2 , Michael Ryckelynck 1
Nature Chemical Biology ( IF 12.9 ) Pub Date : 2019-10-21 , DOI: 10.1038/s41589-019-0381-8 Farah Bouhedda 1 , Kyong Tkhe Fam 2 , Mayeul Collot 2 , Alexis Autour 1 , Stefano Marzi 1 , Andrey Klymchenko 2 , Michael Ryckelynck 1
Affiliation
Live-cell imaging of RNA has remained a challenge because of the lack of naturally fluorescent RNAs. Recently developed RNA aptamers that can light-up small fluorogenic dyes could overcome this limitation, but they still suffer from poor brightness and photostability. Here, we propose the concept of a cell-permeable fluorogenic dimer of self-quenched sulforhodamine B dyes (Gemini-561) and the corresponding dimerized aptamer (o-Coral) that can drastically enhance performance of the current RNA imaging method. The improved brightness and photostability, together with high affinity of this complex, allowed direct fluorescence imaging in live mammalian cells of RNA polymerase III transcription products as well as messenger RNAs labeled with a single copy of the aptamer; that is, without tag multimerization. The developed fluorogenic module enables fast and sensitive detection of RNA inside live cells, while the proposed design concept opens the route to new generation of ultrabright RNA probes.
中文翻译:
用于活细胞 RNA 成像的基于二聚化的荧光染料适配体模块。
由于缺乏天然荧光 RNA,RNA 的活细胞成像仍然是一个挑战。最近开发的可以点亮小荧光染料的 RNA 适配体可以克服这一限制,但它们仍然存在亮度和光稳定性差的问题。在这里,我们提出了自淬灭硫罗丹明 B 染料 (Gemini-561) 和相应的二聚化适体 (o-Coral) 的细胞渗透性荧光二聚体的概念,可以显着提高当前 RNA 成像方法的性能。提高的亮度和光稳定性,以及这种复合物的高亲和力,允许在活的哺乳动物细胞中对 RNA 聚合酶 III 转录产物以及标记有单个适体拷贝的信使 RNA 进行直接荧光成像;也就是说,没有标签多聚化。
更新日期:2019-10-21
中文翻译:
用于活细胞 RNA 成像的基于二聚化的荧光染料适配体模块。
由于缺乏天然荧光 RNA,RNA 的活细胞成像仍然是一个挑战。最近开发的可以点亮小荧光染料的 RNA 适配体可以克服这一限制,但它们仍然存在亮度和光稳定性差的问题。在这里,我们提出了自淬灭硫罗丹明 B 染料 (Gemini-561) 和相应的二聚化适体 (o-Coral) 的细胞渗透性荧光二聚体的概念,可以显着提高当前 RNA 成像方法的性能。提高的亮度和光稳定性,以及这种复合物的高亲和力,允许在活的哺乳动物细胞中对 RNA 聚合酶 III 转录产物以及标记有单个适体拷贝的信使 RNA 进行直接荧光成像;也就是说,没有标签多聚化。