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Fluorescent Visualization of Nucleolar G-Quadruplex RNA and Dynamics of Cytoplasm and Intranuclear Viscosity
CCS Chemistry ( IF 11.2 ) Pub Date : 2020-12-24 , DOI: 10.31635/ccschem.020.202000479
Le Yu 1, 2 , Peter Verwilst 3 , Inseob Shim 2 , Yu-Qiang Zhao 1 , Ying Zhou 1 , Jong Seung Kim 2
Affiliation  

The nucleolus, the locus of ribosome biogenesis, was found to be the predominant intracellular target of a new fluorescent probe, V-P1. In solution, the probe demonstrated both a selectivity to RNA G-quadruplexes and a sensitivity to the viscosity, while G-quadruplex binding did not disturb the viscosity sensing. In cells, confocal and fluorescence lifetime imaging, combined with digestion and competition experiments, lent support to the hypothesis of an RNA-based G-quadruplex as the intracellular target, postulated to be nucleolar ribosomal RNA (rRNA). The probe demonstrated a high sensitivity to viscosity in both the cytoplasm and the nuclear compartment and was used to precisely interrogate the viscosity changes resulting from diverse stimuli, such as temperature, monensin treatment, and etoposide-induced apoptosis. Owing to the putative rRNA G-quadruplex binding in vitro and in vivo, and further combined with a relatively low degree of toxicity, the dye enabled the interrogation of cytoplasm and intranuclear viscosity changes under diverse conditions and found applications in studying the influence and significance of cytoplasm and intranuclear viscosity as well as in gaining insight into the native secondary structure of rRNA in nucleoli.



中文翻译:

荧光可视化的核G四联体RNA和细胞质和核内粘度的动力学。

发现核仁是核糖体生物发生的场所,是新荧光探针的主要细胞内靶标, V一1。在溶液中,该探针显示出对RNA G-四链体的选择性和对粘度的敏感性,而G-四链体结合不会干扰粘度感测。在细胞中,共聚焦和荧光寿命成像,与消化和竞争实验相结合,为基于RNA的G-四链体作为细胞内靶标的假说提供了支持,假定其为核仁核糖体RNA(rRNA)。该探针对细胞质和细胞核室中的粘度都表现出很高的敏感性,并用于精确询问由多种刺激(例如温度,莫能菌素处理和依托泊苷诱导的细胞凋亡)引起的粘度变化。由于假定的rRNA G-四链体在体内和体外结合,并进一步结合了较低的毒性,

更新日期:2020-12-25
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