Qu Chun-Yan, Zhou Li-Sha, Shu Li-Hong, Huang Qian
Laboratory Department, The Seventh People's Hospital of Chengdu, Chengdu 610021, China.
ACS Omega. 2021 Jan 20;6(4):3330-3335. doi: 10.1021/acsomega.0c05798. eCollection 2021 Feb 2.
MicroRNAs (miRNAs) modulate a variety of cellular signaling pathways and play a vital role in cell-to-cell communication. The overlapped expression of a certain miRNA is commonly reported to be related to cancers. Therefore, combined detection of multiple miRNAs is of great significance for cancer diagnosis. Herein, we developed a Fe L tetrahedron-assisted three-way junction (3WJ) probe, which exhibited a higher stability than the normal 3WJ probe, for multiple miRNA detection. In this method, the simultaneous existence of miRNA-21 and miRNA-144 triggers the release of the Y3 sequence in the Fe L tetrahedron-assisted 3WJ probe, which in turn triggers subsequent CRISPR-Cas12a-assisted rolling circle amplification. Based on this, simultaneous detection of miRNA-21 and miRNA-144 was achieved. Furthermore, we also applied this method to the detection of miRNAs in clinical samples and achieved good agreement with quantitative real-time polymerase chain reaction (qRT-PCR), indicating its significant potentials in early diagnosis and treatment of cancer.
微小RNA（miRNA）可调节多种细胞信号通路，并在细胞间通讯中发挥至关重要的作用。据普遍报道，某种miRNA的重叠表达与癌症相关。因此，联合检测多种miRNA对癌症诊断具有重要意义。在此，我们开发了一种铁（Fe）L四面体辅助的三向接头（3WJ）探针，用于多种miRNA检测，该探针表现出比普通3WJ探针更高的稳定性。在该方法中，miRNA-21和miRNA-144的同时存在会触发铁L四面体辅助的3WJ探针中Y3序列的释放，进而触发随后的CRISPR-Cas12a辅助滚环扩增。基于此，实现了对miRNA-21和miRNA-144的同时检测。此外，我们还将该方法应用于临床样本中miRNA的检测，并与定量实时聚合酶链反应（qRT-PCR）取得了良好的一致性，表明其在癌症早期诊断和治疗中具有显著潜力。
