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Related Experiment Video

Updated: May 15, 2026

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors
06:07

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors

Published on: August 5, 2022

Microarray-based fluorescence assay of endonuclease functionality and inhibition.

Lan Ma1, Min Su, Tao Li

  • 1State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, PR China.

The Analyst
|January 4, 2013
PubMed
Summary
This summary is machine-generated.

A novel double-strand DNA microarray assay detects endonuclease activity and inhibition. This method shows high specificity for enzymes like EcoRI and BamHI, enabling high-throughput screening of potential inhibitors.

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Published on: December 23, 2013

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Last Updated: May 15, 2026

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06:10

A Fluorescence-based Exonuclease Assay to Characterize DmWRNexo, Orthologue of Human Progeroid WRN Exonuclease, and Its Application to Other Nucleases

Published on: December 23, 2013

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Assay Development

Background:

  • Endonucleases play crucial roles in DNA metabolism and are targets for therapeutic intervention.
  • Developing sensitive and specific assays for endonuclease activity and inhibition is essential for drug discovery and molecular diagnostics.

Purpose of the Study:

  • To develop and validate a double-strand (ds) DNA microarray-based fluorescence assay for studying endonuclease functionality.
  • To assess the potential of this assay for high-throughput screening of endonuclease inhibitors.

Main Methods:

  • Fabrication of dsDNA microarrays by hybridizing Cy5-labeled oligonucleotides with immobilized complementary probes.
  • Utilizing fluorescence decrease as an indicator of endonuclease-mediated dsDNA cleavage.
  • Testing the assay with two endonucleases (EcoRI, BamHI) and four potential inhibitors (doxorubicin hydrochloride, 5-fluorouracil, ethidium bromide, actinomycin D).

Main Results:

  • The assay demonstrated significant fluorescence decrease upon endonuclease activity.
  • High specificity was achieved, with detection limits of 1.1 U/mL for EcoRI and 2.0 U/mL for BamHI.
  • The assay successfully detected inhibition of EcoRI and BamHI by the tested compounds, proving its utility for inhibitor screening.

Conclusions:

  • A robust dsDNA microarray fluorescence assay for endonuclease activity and inhibition has been successfully developed.
  • The assay exhibits high sensitivity and specificity, suitable for detecting enzyme function.
  • This platform holds significant potential for high-throughput screening of endonuclease inhibitors in drug discovery efforts.