A facile deoxyuridine/biotin-modified molecular beacon for simultaneous detection of proteins and nucleic acids via a label-free and background-eliminated fluorescence assay

The Analyst
Fei YinChenzhong Li

Abstract

Simultaneous detection of different types of cancer biomarkers (nucleic acids and proteins) could facilitate early diagnosis of cancer and clinical treatment. Herein, a simultaneous detection platform of proteins and nucleic acids has been developed using a single substrate probe combining a label-free and background-eliminated fluorescence assay. Telomerase and telomerase RNA (TR) were chosen as the models. The molecular beacon (dU-BIO-HP) that contains deoxyuridine/biotin in its side arm, a TR recognition sequence in the loop and a telomerase substrate primer at the stem end was ingeniously designed. In the presence of telomerase, the stem of dU-BIO-HP is elongated by the addition of telomere repeats complementary to the assistant DNA. Furthermore, the formed dsDNA performed as engaging primers to initiate a SDA reaction, generating abundant G-quadruplex monomers. Similarly, on TR, the hybridization between TR and dU-BIO-HP can open its stem, triggering another SDA reaction, producing abundant short ssDNAs. With the G-quadruplex binding with ZnPPIX and ssDNA binding with SG for specific fluorescence responses, the label-free multiple detection can be achieved. In our strategy, the deoxyuridine of dU-BIO-HP acts as a barrier t...Continue Reading

References

Jul 26, 1996·The Journal of Biological Chemistry·R S LaskenA Rashtchian
Nov 20, 2012·Chemical Communications : Chem Comm·Tian TianXiang Zhou
Mar 9, 2013·Journal of Electroanalytical Chemistry·Subbiah AlwarappanChen-Zhong Li
Oct 29, 2013·Chemical Society Reviews·Jinbo LiYan Zhang
Mar 13, 2014·Expert Review of Proteomics·Haitham A BadrChen-Zhong Li
Jun 22, 2017·ACS Applied Materials & Interfaces·Ruixue DuanFan Xia
Aug 2, 2017·Journal of the American Chemical Society·Wei MaChuanlai Xu
Oct 25, 2017·Analytical Chemistry·Haitang YangSongqin Liu
Oct 4, 2018·Analytica Chimica Acta·Li ZouGongke Li

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Methods Mentioned

BETA
Fluorescence
electrophoresis
fluorescence assay

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