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Updated: Sep 8, 2025
08:56
A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
Published on: December 5, 2016
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Detecting and Mapping N6-Methyladenosine on RNA/DNA Hybrids
Abdulkadir Abakir1, Fahad Alenezi2, Alexey Ruzov3
1Epigenetic Programme, Babraham Institute, Cambridge, UK. abdulkadir.abakir@babraham.ac.uk.
Methods in Molecular Biology (Clifton, N.J.)
|June 15, 2022
Summary
N6-methyladenosine (m6A) modifies RNA in R-loops, impacting gene expression and genome stability. New methods detect and map these m6A-decorated RNA/DNA hybrids in cells.
Area of Science:
- Molecular Biology
- Epigenetics
- Genetics
Background:
- N6-methyladenosine (m6A) is a crucial RNA modification for gene expression regulation in eukaryotes.
- R-loops, composed of RNA/DNA hybrids and a DNA strand, are implicated in genetic instability and gene regulation.
- m6A modification of RNA within R-loops influences their stability and metabolism.
Purpose of the Study:
- To present novel methodologies for detecting and analyzing m6A-modified RNA/DNA hybrids.
- To enable investigation of the spatial distribution and genomic mapping of these structures.
Main Methods:
- Immunostaining protocol for visualizing m6A-containing RNA/DNA hybrids within cells.
- m6A-DNA immunoprecipitation (DIP) for genome-wide mapping and locus-specific analysis.
Main Results:
- The study introduces two robust methods for R-loop analysis.
- These techniques allow for the detection and spatial and genomic characterization of m6A-modified R-loops.
Conclusions:
- The developed methods facilitate the study of m6A in R-loop biology.
- These techniques can be extended to investigate other RNA modifications in R-loops, advancing our understanding of genome regulation and stability.
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