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Stable Converter-Powered Cascade Amplification of DNAzyme and Cas12a for Electrochemiluminescent Detection of DNA Adenine Methyltransferase
Using a Stable Converter to Boost DNAzyme and Cas12a for Sensitive Light-Based Detection of DNA Adenine Methyltransferase
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Abstract
The proposed biosensor allows for highly accurate and sensitive electrochemiluminescent detection of DNA adenine methyltransferase (Dam) with a detection limit down to 8.61 × 10⁻⁷ U mL⁻¹.
- A dual-sites dumbbell-shaped converter (DDC) was developed for reliable signal conversion in detecting Dam activity.
- The DDC structure is methylated by Dam and cleaved by DpnI, converting Dam activity into a mimic target (MT).
- Hybridization of MT with a smart allosteric spherical DNAzyme (SASD) activates a DNAzyme that releases a Cas12a activator.
- The activated Cas12a cleaves quenching probes on the electrochemiluminescent electrode, restoring the ECL signal for quantitative analysis.
- The biosensor demonstrates a wide linear detection range from 1.0 × 10⁻⁶ to 1.0 U mL⁻¹.
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