School of Chemistry and Biology Engineering, University of Science and Technology Beijing, Beijing, 100083, P. R. China.
Org Biomol Chem. 2020 May 6;18(17):3269-3273. doi: 10.1039/d0ob00625d.
Circular RNAs (circRNAs), which function as a kind of newly discovered biomarker, have been shown to play an important role in gene expression, signaling pathway and disease diagnosis. Accurate recognition and sensitive detection of sequence-specific circRNA of interest can advance our understanding of its biological functions and the diagnosis of human diseases. Herein, we have newly developed a highly sensitive and specific method to quantitatively detect circRNA through accurate ligation of two ingeniously designed DNA probes using splintR ligase at the junction site of circRNA, which can directly discriminate circRNA from the corresponding linear RNA. With PCR amplification of the ligated DNA probes, circRNA as low as 1 fM can be detected with the dynamic range spanning over five orders of magnitude. This method has been successfully applied to the detection of circRNA in total RNA samples extracted from cell lines.
环状 RNA（circRNAs）作为一种新发现的生物标志物，已被证明在基因表达、信号通路和疾病诊断中发挥着重要作用。准确识别和敏感检测感兴趣的序列特异性 circRNA 可以促进我们对其生物学功能和人类疾病诊断的理解。在这里，我们开发了一种新的方法，通过在 circRNA 连接点处使用 splintR 连接酶精确连接两个精心设计的 DNA 探针，来定量检测 circRNA，该方法可以直接区分 circRNA 和相应的线性 RNA。通过连接 DNA 探针的 PCR 扩增，可以检测低至 1 fM 的 circRNA，其动态范围跨越五个数量级。该方法已成功应用于从细胞系提取的总 RNA 样品中 circRNA 的检测。
