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Updated: May 26, 2026
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Continuous Fluorescence-Based Endonuclease-Coupled DNA Methylation Assay to Screen for DNA Methyltransferase Inhibitors
Published on: August 5, 2022
Simultaneous single-molecule detection of endogenous C-5 DNA methylation and chromatin accessibility using MAPit
Russell P Darst1, Carolina E Pardo, Santhi Pondugula
1Department of Biochemistry and Molecular Biology, University of Florida and Shands Cancer Center, University of Florida College of Medicine, Gainesville, FL, USA.
Methods in Molecular Biology (Clifton, N.J.)
|December 21, 2011
Summary
Methylation accessibility protocol for individual templates (MAPit) reveals DNA accessibility and protein binding sites. This technique uses exogenous methylation to map genome regions, enhancing single-molecule DNA methylation analysis.
Area of Science:
- Epigenetics
- Genomics
- Molecular Biology
Background:
- Bisulfite sequencing offers single-molecule DNA methylation analysis.
- Current methods have limitations in mapping genome accessibility.
Purpose of the Study:
- To develop a novel technique for mapping DNA accessibility and protein binding sites.
- To enhance single-molecule DNA methylation analysis by incorporating exogenous methylation information.
Main Methods:
- Developed Methylation Accessibility Protocol for Individual Templates (MAPit).
- Utilized exogenous DNA methyltransferase (DNMT) treatment prior to DNA extraction.
- Combined endogenous cytosine methylation analysis with exogenous GC methylation mapping.
Main Results:
- MAPit successfully maps regions of DNA accessibility.
- Exogenous methylation footprints nucleosomes and other DNA-binding proteins.
- Each sequenced molecule provides a record of both endogenous and exogenous methylation states.
Conclusions:
- MAPit provides a powerful tool for understanding genome accessibility at single-molecule resolution.
- The technique integrates DNA methylation analysis with protein-DNA interaction mapping.
- MAPit significantly advances the capabilities of genomic and epigenomic research.
