Liang Huiping, Mu Xiaomei, Huang Yong, Zhao Shulin, Tian Jianniao
State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources, Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), School of Chemistry and Pharmaceutical Sciences, Guangxi Normal University, Guilin, 541004, China.
Chem Asian J. 2025 Mar 3;20(5):e202401209. doi: 10.1002/asia.202401209. Epub 2025 Jan 15.
The anomalous expression of microRNA poses a serious threat to human life and health safety, and serves as an important biomarker for cancer detection. In this study, a novel magnetic-assisted DNA logic gate nanomachine triggered by miRNA-21 and miRNA-155 was designed based on the trans-cleavage activity of CRISPR/Cas12a activated by a split DNA activator, using only a single crRNA and signal probe, which simplified the detection procedure and complex nucleic acid amplification. The presence of target molecules, miRNA-21 and miRNA-155, can stimulate the DNA walker machine assembled on magnetic beads, which releases activator under the action of DNAzyme. Then the trans-cleavage activity of CRISPR/Cas12a is initiated and the system signal significantly increases. Based on this, an AND logic gate nanomachine was constructed for simultaneous analysis of miRNA-21 and miRNA-155. The detection limits of miRNA-21 and miRNA-15 were 9.00 pM and 42.00 pM, respectively, and this method was successfully applied to miRNA analysis in cell samples. This nanomachine combined the DNA walker with DNA logic circuit and CRISPR/Cas12a system, providing a new approach for simultaneous detection of multiple targets and further expanding the application of gene editing in the analysis and sensing of multiple target substances.
微小RNA的异常表达对人类生命和健康安全构成严重威胁，并且是癌症检测的重要生物标志物。在本研究中，基于由分裂DNA激活剂激活的CRISPR/Cas12a的反式切割活性，设计了一种由miRNA-21和miRNA-155触发的新型磁辅助DNA逻辑门纳米机器，仅使用单个crRNA和信号探针，简化了检测程序和复杂的核酸扩增。靶分子miRNA-21和miRNA-155的存在可以刺激组装在磁珠上的DNA步行机，其在脱氧核酶的作用下释放激活剂。然后启动CRISPR/Cas12a的反式切割活性，系统信号显著增加。基于此，构建了一个与门逻辑门纳米机器用于同时分析miRNA-21和miRNA-155。miRNA-21和miRNA-15的检测限分别为9.00 pM和42.00 pM，该方法已成功应用于细胞样品中的miRNA分析。这种纳米机器将DNA步行机与DNA逻辑电路和CRISPR/Cas12a系统相结合，为同时检测多个靶标提供了一种新方法，并进一步扩展了基因编辑在多种靶标物质分析和传感中的应用。
