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Related Experiment Video

Updated: May 7, 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

Genetic sensor for strong methylating compounds.

Felix Moser, Andrew Horwitz, Jacinto Chen

    ACS Synthetic Biology
    |September 17, 2013
    PubMed
    Summary
    This summary is machine-generated.

    Researchers engineered a bacterial DNA repair protein, Ada, into a sensitive yeast sensor for toxic methylating chemicals. This new sensor effectively detects environmental pollutants at low concentrations.

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    Correlating Gene-specific DNA Methylation Changes with Expression and Transcriptional Activity of Astrocytic KCNJ10 (Kir4.1)

    Published on: September 26, 2015

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Environmental Science

    Background:

    • Methylating chemicals are prevalent in industry and agriculture, posing toxicity risks due to DNA methylation.
    • The Escherichia coli Ada protein naturally senses methyl adducts on DNA's phosphoester backbone.

    Purpose of the Study:

    • To characterize the E. coli Ada protein as a genetic sensor for methylating agents.
    • To engineer a more sensitive sensor by lowering its detection threshold.
    • To develop a functional eukaryotic sensor for methyl phosphotriester adducts.

    Main Methods:

    • Overexpression of Ada in E. coli to enhance dynamic range and lower detection threshold.
    • Fusion of the N-terminal domain of Ada with the Gal4 transcriptional activation domain in Saccharomyces cerevisiae.
    • Tuning sensor specifications by altering expression levels and Ada operator numbers.

    Main Results:

    • Enhanced E. coli Ada sensor achieved 350-fold induction and a 40 microM detection threshold for methyl iodide.
    • Developed a functional yeast sensor for methyl phosphotriester adducts with a 28 microM detection threshold and 5.2-fold induction.
    • The yeast sensor exhibited a response profile similar to native Ada, indicating retained function across species.

    Conclusions:

    • The engineered yeast sensor is suitable for detecting relevant environmental concentrations of methylating compounds.
    • This work successfully transferred a prokaryotic sensor to a eukaryotic system with rational tuning.
    • The developed sensor provides a novel tool for monitoring environmental exposure to toxic methylating agents.