Asan Institute for Life Science, Asan Medical Center, Seoul 05505, Korea.
Department of Convergence Medicine, University of Ulsan College of Medicine, Seoul 05505, Korea.
Int J Mol Sci. 2021 Jun 16;22(12):6433. doi: 10.3390/ijms22126433.
Ribonucleic acid (RNA) plays an important role in many cellular processes. Thus, visualizing and quantifying the molecular dynamics of RNA directly in living cells is essential to uncovering their role in RNA metabolism. Among the wide variety of fluorescent probes available for RNA visualization, exciton-controlled hybridization-sensitive fluorescent oligonucleotide (ECHO) probes are useful because of their low fluorescence background. In this study, we apply fluorescence correlation methods to ECHO probes targeting the poly(A) tail of mRNA. In this way, we demonstrate not only the visualization but also the quantification of the interaction between the probe and the target, as well as of the change in the fluorescence brightness and the diffusion coefficient caused by the binding. In particular, the uptake of ECHO probes to detect mRNA is demonstrated in HeLa cells. These results are expected to provide new insights that help us better understand the metabolism of intracellular mRNA.
核糖核酸 (RNA) 在许多细胞过程中发挥着重要作用。因此,直接在活细胞中可视化和定量 RNA 的分子动力学对于揭示其在 RNA 代谢中的作用至关重要。在可用于 RNA 可视化的各种荧光探针中,由于背景荧光低,激子控制杂交敏感荧光寡核苷酸 (ECHO) 探针非常有用。在这项研究中,我们应用荧光相关方法研究靶向 mRNA 聚 (A) 尾的 ECHO 探针。通过这种方式,我们不仅证明了探针与靶标之间相互作用的可视化,还证明了结合引起的荧光强度和扩散系数变化的定量,以及 ECHO 探针用于检测 mRNA 的摄取。这些结果有望提供新的见解,帮助我们更好地理解细胞内 mRNA 的代谢。