Institute of Organic Chemistry and Chemical Biology, Goethe-University Frankfurt, Max-von-Laue Str. 7-9, 60438, Frankfurt, Germany.
Institute of Physical and Theoretical Chemistry, Goethe-University Frankfurt, Max-von-Laue Str. 7-9, 60438, Frankfurt, Germany.
Chem Asian J. 2022 Dec 14;17(24):e202201077. doi: 10.1002/asia.202201077. Epub 2022 Nov 18.
Protein synthesis via ribosomes is a fundamental process in all known living organisms. However, it can be completely stalled by removing a single nucleobase (depurination) at the sarcin/ricin loop of the ribosomal RNA. In this work, we describe the preparation and optimization process of a fluorescent probe that can be used to visualize depurination. Starting from a fluorescent thiophene nucleobase analog, various RNA probes that fluoresce exclusively in the presence of a depurinated sarcin/ricin-loop RNA were designed and characterized. The main challenge in this process was to obtain a high fluorescence signal in the hybridized state with an abasic RNA strand, while keeping the background fluorescence low. With our new RNA probes, the fluorescence intensity and lifetime can be used for efficient monitoring of depurinated RNA.
核糖体介导的蛋白质合成是所有已知生物的基本过程。然而，通过从核糖体 RNA 的 Sarcin/ricin 环中去除单个核苷（去嘌呤）可以完全阻止该过程。在这项工作中，我们描述了一种荧光探针的制备和优化过程，该探针可用于可视化去嘌呤化。从荧光噻吩核苷类似物出发，设计并表征了各种仅在存在去嘌呤化 Sarcin/ricin 环 RNA 时才发出荧光的 RNA 探针。在此过程中的主要挑战是在与无碱基 RNA 链杂交时获得高荧光信号，同时保持低背景荧光。使用我们的新 RNA 探针，可以有效地监测去嘌呤化 RNA 的荧光强度和寿命。
