Wang Rui, Wang Shuai, Xu Xiaowen, Jiang Wei, Zhang Nan
Shandong Key Laboratory of Biophysics, Institute of Biophysics, Dezhou University, 253023, Dezhou, PR China; School of Chemistry and Chemical Engineering, Shandong University, 250100, Jinan, PR China.
School of Chemistry and Chemical Engineering, Shandong University, 250100, Jinan, PR China.
Anal Chim Acta. 2021 Mar 8;1149:338213. doi: 10.1016/j.aca.2021.338213. Epub 2021 Jan 11.
Here, a MNAzyme probes mediated DNA logic platform was developed for microRNAs (miRNAs) logic detection and cancer cells identification. A series of MNAzyme probes containing the cleavage active center were designed. Four types of logic gates were constructed, including YES, AND, XOR and NOR gate. These logic gates used miRNAs that were high expression in cancer cells as logic inputs and used MNAzyme cleavage amplification reaction to output signals. For the construction of intracellular logic gates, MnO nanosheets were used as carriers and cofactor providers. When MnO nanoprobes entered the cells through endocytosis, the intracellular glutathione degraded the MnO nanosheets to release the cofactor Mn and MNAzyme probes. The MNAzyme probes bound to the miRNAs and catalyze the MNAzyme cleavage amplification reaction, producing enhanced fluorescent signal with "true" output. The logic detection of miRNAs was achieved by integrating information from the AND, XOR and NOR logic gates. Moreover, through the construction of intracellular YES and AND logic gates, the cancer cells identification, especially the identification of same type of cancer cells with different phenotypes was achieved. These experimental results showed that this platform held great promise in accurate diagnosis and treatment of cancer.
在此,开发了一种基于MNAzyme探针的DNA逻辑平台,用于微小RNA(miRNA)的逻辑检测和癌细胞识别。设计了一系列含有切割活性中心的MNAzyme探针。构建了四种类型的逻辑门,包括“是”门、“与”门、“异或”门和“或非”门。这些逻辑门使用癌细胞中高表达的miRNA作为逻辑输入,并利用MNAzyme切割扩增反应输出信号。为构建细胞内逻辑门,MnO纳米片被用作载体和辅因子提供者。当MnO纳米探针通过内吞作用进入细胞时,细胞内的谷胱甘肽降解MnO纳米片以释放辅因子Mn和MNAzyme探针。MNAzyme探针与miRNA结合并催化MNAzyme切割扩增反应,产生增强的荧光信号并输出“真”值。通过整合“与”门、“异或”门和“或非”门的信息实现了miRNA的逻辑检测。此外,通过构建细胞内“是”门和“与”门,实现了癌细胞识别,特别是对具有不同表型的同类型癌细胞的识别。这些实验结果表明,该平台在癌症的准确诊断和治疗方面具有巨大潜力。