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Epigenetic Regulation

Epigenetic Regulation

Epigenetic changes alter the physical structure of the DNA without changing the genetic sequence and often regulate whether genes are turned on or off. This regulation ensures that each cell produces only proteins necessary for its function. For example, proteins that promote bone growth are not produced in muscle cells. Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.
X-chromosome...

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Related Experiment Video

Updated: Sep 29, 2025

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

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

Published on: August 5, 2022

Profiling demethylase activity using epigenetically inactivated DNAzyme.

Juan Huang¹, Jiao Wang¹, Zhenkun Wu¹

¹State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biomedical Sciences, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, China.

Biosensors & Bioelectronics

|March 22, 2022

Summary

This summary is machine-generated.

This study introduces a novel assay to measure active demethylase levels, crucial for understanding cancer chemoresistance. The new method accurately quantifies O6-methylguanine-DNA-methyltransferase (MGMT) activity, offering a sensitive tool for disease research.

Keywords:

CRISPR/Cas12a DNAzyme Demethylase Epigenetic modification

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Last Updated: Sep 29, 2025

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Area of Science:

Biochemistry
Molecular Biology
Cancer Research

Background:

DNA methylation is linked to various carcinomas and regulated by demethylases.
Current antibody-based methods measure demethylase concentration, not active enzyme levels.
Understanding active demethylase levels is critical for cancer therapy and chemoresistance.

Purpose of the Study:

To develop a novel, ultrasensitive activity assay for disease-related demethylases.
To quantify the catalytic activity of O6-methylguanine-DNA-methyltransferase (MGMT).
To establish a new method for profiling active demethylase levels in disease contexts.

Main Methods:

Development of an activity assay using epigenetically modified DNAzymes (EMOzymes).
Utilized CRISPR/Cas12a facilitated cascade signal amplification for enhanced sensitivity.
Quantified the activity of O6-methylguanine-DNA-methyltransferase (MGMT).

Main Results:

Achieved ultrasensitive quantification of MGMT activity.
Established a limit of detection as low as 0.054 nM for MGMT activity.
Demonstrated the assay's potential for profiling various disease-related demethylases.

Conclusions:

The developed EMOzyme and CRISPR/Cas12a-based assay provides a sensitive method for measuring active demethylase levels.
This approach overcomes limitations of antibody-based assays by quantifying enzyme activity.
Opens new avenues for understanding the role of demethylases in cancer and developing targeted therapies.