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Related Experiment Video

Updated: Mar 27, 2026

Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors
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Reduced Maintenance DNA Methylation Thresholds Enable Sensitive Reporter Assays for UHRF1 and DNMT1 Inhibition.

Cuicui Xia1,2, Ying Cui3, Jiongliang Wang2

  • 1Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China.

Advanced Science (Weinheim, Baden-Wurttemberg, Germany)
|March 26, 2026
PubMed
Summary

Discovering new cancer drugs is challenging. This study reveals that targeting UHRF1 and DNMT1, key proteins in DNA methylation, can overcome drug resistance and improve cancer therapies.

Keywords:
DNA hypomethylating agentsDNMT1 inhibitionHTS assay developmentUHRF1 inhibitionrecombinant reporter assay

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Area of Science:

  • Epigenetics and Cancer Biology
  • Molecular Oncology
  • Drug Discovery

Background:

  • Aberrant DNA methylation silences tumor suppressor genes (TSGs) via the UHRF1-DNMT1 axis.
  • Current hypomethylating agents (HMAs) face challenges due to stringent thresholds for DNMT1 inhibition.
  • UHRF1 also exhibits stringent thresholds in maintaining TSG suppression.

Purpose of the Study:

  • To investigate the role of maintenance methylation thresholds in TSG suppression.
  • To develop sensitive reporter systems for monitoring HMA-induced TSG reactivation.
  • To facilitate the discovery of novel HMAs and epigenetic combination therapies.

Main Methods:

  • Genetically reduced redundancy in DNMT1 or UHRF1 to lower inhibition thresholds.
  • Engineered recombinant reporter systems linked to endogenous hypermethylated promoters in hypomorphic cells.
  • Pilot high-throughput screening of DNMT1 and UHRF1 inhibitors.

Main Results:

  • Reducing DNMT1 or UHRF1 redundancy significantly lowered inhibition thresholds and sensitized cells.
  • Reporter systems demonstrated improved sensitivity and dynamic range for monitoring TSG reactivation.
  • High-throughput screening validated the robustness and specificity of the developed assays.

Conclusions:

  • Maintenance methylation thresholds are critical in TSG suppression.
  • The study provides versatile tools for discovering novel HMAs.
  • Developed platforms enable optimization of epigenetic combination therapies for enhanced TSG re-expression.