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Chem Sci. 2018 Nov 28;10(6):1709-1715. doi: 10.1039/c8sc03305f. eCollection 2019 Feb 14.

Sensitively distinguishing intracellular precursor and mature microRNA abundance.

Yang F1,2, Cheng Y1,2, Cao Y1,2, Dong H1,2, Lu H1,2, Zhang K1,2, Meng X1,2, Liu C1,2, Zhang X1,2.

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Beijing Advanced Innovation Center for Materials Genome Engineering , University of Science and Technology Beijing , 30 Xueyuan Road , Beijing 100083 , P. R. China . Email: ; Email:
Beijing Key Laboratory for Bioengineering and Sensing Technology , Research Center for Bioengineering and Sensing Technology , School of Chemistry & Biological Engineering , University of Science & Technology Beijing , 30 Xueyuan Road , Beijing 100083 , P. R. China.


Mature microRNAs (miRNAs) produced from precursor microRNAs (pre-miRNAs) by the RNase Dicer have showed significant potential for cancer diagnosis and prognosis due to their key regulatory roles in various pathological processes. However, discriminatory detection of low-abundance miRNAs and pre-miRNAs remains a key challenge since the mature sequence is also present in the pre-miRNA forms. Herein, we report a novel cascade reaction to sensitively distinguish miRNAs versus pre-miRNAs in living cells based on two pairs of programmable hairpin oligonucleotide probes with a simple sequence design. The programmable hairpin probes can metastably coexist until the introduction of miRNAs or pre-miRNAs, which can trigger a specific hybridization chain reaction (HCR), respectively, leading to the self-assembly of nicked DNA duplex structures and a remarkable specific fluorescence intensity increase. The system can readily and sensitively assess the miRNA or pre-miRNA abundance in a homogeneous solution. The intracellular miRNA and pre-miRNA expression level assessment in different living cells is realized. Thus, we provide a novel investigation tool for discriminatorily and accurately assessing miRNA and pre-miRNA abundance, which could be useful for the biomedical application of miRNAs.

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