基于布尔逻辑运算的端粒酶和 MiR-21 的超灵敏双信号检测,ACS Applied Materials & Interfaces

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基于布尔逻辑运算的端粒酶和 MiR-21 的超灵敏双信号检测
ACS Applied Materials & Interfaces ( IF 8.3 ) Pub Date : 2021-10-19 , DOI: 10.1021/acsami.1c17912
Jiaju Shi ₁ , Meiqi Shen ₁ , Wenjie Zhao ₂ , Jinhua Liu ₂ , Zongjin Qu ₁ , Mengting Zhu ₁ , Zichao Chen ₃ , Pengfei Shi ₁ , Zhen Zhang _{1,

2} , Shu-Sheng Zhang ₁

Affiliation

端粒酶和微 RNA (miRNA) 在多种肿瘤细胞中同时上调，并已成为有前景的肿瘤标志物。然而，由于端粒酶和 miRNA 的低表达水平，原位的灵敏检测仍然是一个巨大的挑战。在这里，我们设计了一种基于功能化有序介孔纳米粒子（FOMN）的布尔逻辑“AND”信号放大策略，以实现对活肿瘤细胞中端粒酶和 miR-21 的超灵敏检测。简而言之，该策略使用端粒酶作为输入，通过使包裹的 DNA1 在 dNTP 的共同参与下形成 DNA-a 发夹，从而释放 DNA3-ROX-BHQ 发夹。随后，DNA2-Ag、DNA3-ROX-BHQ和第二输入miR-21参与杂交链式反应，放大荧光和拉曼信号。实验结果显示输出双信号的强度与端粒酶和miR-21的表达水平相关。Ag 纳米粒子 (AgNPs) 不仅增强了荧光信号，而且还允许获得更灵敏的拉曼信号。因此，即使肿瘤标志物的表达处于低水平，基于FOMN的双信号逻辑运算策略仍然可以实现对端粒酶和miR-21的原位灵敏检测。此外，FOMNs 可以在短时间内检测 miR-21 的表达水平。因此，该策略在检测肿瘤标志物和评估肿瘤治疗效果方面具有潜在的临床应用价值。Ag 纳米粒子 (AgNPs) 不仅增强了荧光信号，而且还允许获得更灵敏的拉曼信号。因此，即使肿瘤标志物的表达处于低水平，基于FOMN的双信号逻辑运算策略仍然可以实现对端粒酶和miR-21的原位灵敏检测。此外，FOMNs 可以在短时间内检测 miR-21 的表达水平。因此，该策略在检测肿瘤标志物和评估肿瘤治疗效果方面具有潜在的临床应用价值。Ag 纳米粒子 (AgNPs) 不仅增强了荧光信号，而且还允许获得更灵敏的拉曼信号。因此，即使肿瘤标志物的表达处于低水平，基于FOMN的双信号逻辑运算策略仍然可以实现对端粒酶和miR-21的原位灵敏检测。此外，FOMNs 可以在短时间内检测 miR-21 的表达水平。因此，该策略在检测肿瘤标志物和评估肿瘤治疗效果方面具有潜在的临床应用价值。

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Ultrasensitive Dual-Signal Detection of Telomerase and MiR-21 Based on Boolean Logic Operations

Telomerase and micro-RNAs (miRNAs) are simultaneously upregulated in a variety of tumor cells and have emerged as promising tumor markers. However, sensitive detection of telomerase and miRNAs in situ remains a great challenge due to their low expression levels. Here, we designed a Boolean logic “AND” signal amplification strategy based on functionalized ordered mesoporous nanoparticles (FOMNs) to achieve ultrasensitive detection of telomerase and miR-21 in living tumor cells. Briefly, the strategy uses telomerase as an input to enable the release of DNA3-ROX-BHQ hairpins by making the wrapping DNA1 form a DNA-a hairpin with the joint participation of dNTPs. Subsequently, DNA2-Ag, DNA3-ROX-BHQ, and the second input miR-21 participated in hybridization chain reaction to amplify fluorescence and Raman signals. Experimental results showed the intensity of output dual signals relevant to the expression levels of telomerase and miR-21. The Ag nanoparticles (AgNPs) not only enhanced the fluorescence signals but also allowed to obtain more sensitive Raman signals. Therefore, even if expression of tumor markers is at a low level, the FOMN-based dual-signal logic operation strategy can still achieve sensitive detection of telomerase and miR-21 in situ. Furthermore, FOMNs can detect miR-21 expression levels in a short time. Consequently, this strategy has a potential clinical application value in detection of tumor markers and the assessment of tumor treatment efficacy.

更新日期：2021-11-03

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