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.