CRISPR/Cas12a-mediated DNAzyme/split-aptamer cascade for label-free detection of site-specific DNA methylation

Jun 24, 2025Biosensors & bioelectronics

Using CRISPR and DNA tools to detect specific DNA methylation without labels

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Abstract

The assay achieved a linear detection range from 2 to 200 nM with a limit of detection of 1.74 nM.

  • DNA methylation is identified as a significant biomarker for early disease screening.
  • A novel, cascaded, and label-free CRISPR/Cas12a system was developed for detecting Septin9 methylation.
  • Methylation-sensitive digestion was used to selectively target non-methylated DNA, preserving methylated fragments.
  • The method demonstrated accuracy, precision, and selectivity in detecting methylated Septin9 in various sample types.
  • Preliminary results suggest potential applicability in colorectal cancer screening using this detection approach.

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