Yang Fan, Cheng Yaru, Cao Yu, Dong Haifeng, Lu Huiting, Zhang Kai, Meng Xiangdan, Liu Conghui, Zhang Xueji
Beijing Advanced Innovation Center for Materials Genome Engineering , University of Science and Technology Beijing , 30 Xueyuan Road , Beijing 100083 , P. R. China . Email:
Beijing Key Laboratory for Bioengineering and Sensing Technology , Research Center for Bioengineering and Sensing Technology , School of Chemistry & Biological Engineering , University of Science & Technology Beijing , 30 Xueyuan Road , Beijing 100083 , P. R. China.
Chem Sci. 2018 Nov 28;10(6):1709-1715. doi: 10.1039/c8sc03305f. eCollection 2019 Feb 14.
Mature microRNAs (miRNAs) produced from precursor microRNAs (pre-miRNAs) by the RNase Dicer have showed significant potential for cancer diagnosis and prognosis due to their key regulatory roles in various pathological processes. However, discriminatory detection of low-abundance miRNAs and pre-miRNAs remains a key challenge since the mature sequence is also present in the pre-miRNA forms. Herein, we report a novel cascade reaction to sensitively distinguish miRNAs pre-miRNAs in living cells based on two pairs of programmable hairpin oligonucleotide probes with a simple sequence design. The programmable hairpin probes can metastably coexist until the introduction of miRNAs or pre-miRNAs, which can trigger a specific hybridization chain reaction (HCR), respectively, leading to the self-assembly of nicked DNA duplex structures and a remarkable specific fluorescence intensity increase. The system can readily and sensitively assess the miRNA or pre-miRNA abundance in a homogeneous solution. The intracellular miRNA and pre-miRNA expression level assessment in different living cells is realized. Thus, we provide a novel investigation tool for discriminatorily and accurately assessing miRNA and pre-miRNA abundance, which could be useful for the biomedical application of miRNAs.
由核糖核酸酶Dicer从前体微小RNA（pre-miRNA）产生的成熟微小RNA（miRNA），因其在各种病理过程中的关键调控作用，在癌症诊断和预后方面显示出巨大潜力。然而，由于成熟序列也以前体微小RNA的形式存在，低丰度微小RNA和前体微小RNA的鉴别检测仍然是一个关键挑战。在此，我们报道了一种基于两对序列设计简单的可编程发夹寡核苷酸探针，用于在活细胞中灵敏区分微小RNA和前体微小RNA的新型级联反应。可编程发夹探针可以亚稳共存，直到引入微小RNA或前体微小RNA，它们分别触发特定的杂交链式反应（HCR），导致带切口的DNA双链结构自组装，并显著增加特异性荧光强度。该系统可以在均相溶液中轻松灵敏地评估微小RNA或前体微小RNA的丰度。实现了不同活细胞内微小RNA和前体微小RNA表达水平的评估。因此，我们提供了一种用于鉴别和准确评估微小RNA和前体微小RNA丰度的新型研究工具，这可能对微小RNA的生物医学应用有用。
