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Construction of fast-walking tetrahedral DNA walker with efficient electrochemiluminescence for the simultaneous and ultrasensitive detection of m 6 A and m 5 C

Microchemical Journal, ISSN: 0026-265X, Vol: 209, Page: 112611
2025
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Article Description

RNA methylation as a pivotal role in the early diagnosis and prognosis of cancer and associated conditions. This study introduces a new approach for addressing the challenge of ultrasensitive detection of m 6 A-RNA and m 5 C-RNA, offering potential insights into improving diagnostic and prognostic strategies for these diseases. Herein, an endoribonuclease MazF-mediated cleavage reaction was designed to distinguish m 6 A-RNA from m 6 A site-free RNA. Uncleaved m 6 A-RNA (target 1) was efficiently converted to Fc-HP1 by DSN enzyme-assisted amplification to achieve a signal-off state. Subsequently, m 5 C-RNA (target 2) could activate the tetrahedral DNA walker employing Zn 2+ -dependent DNAzyme-specific activation to elicit the “open” ECL response of Ru(bpy) 3 2+ @PtNPs@AuNPs. The findings demonstrated the effectiveness of the “on-off-on” ECL biosensor enabling quantitative detection of m 6 A-RNA with a detection limit of 213 aM and m 5 C-RNA with a detection limit of 2.08 fM. This technique presents an opportunity for the simultaneous detection of epigenetically modified bases in RNA, holding significant promise for the early diagnosis of cancer and other diseases associated with epigenetic modifications.

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