Related Experiment Video
Updated: Apr 5, 2026
08:56
A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
Published on: December 5, 2016
11.4K
Transcriptome-Wide Mapping of N⁶-Methyladenosine by m⁶A-Seq
Dan Dominissini1, Sharon Moshitch-Moshkovitz2, Ninette Amariglio2
1Department of Chemistry, The University of Chicago, Chicago, Illinois, USA; Institute for Biophysical Dynamics, The University of Chicago, Chicago, Illinois, USA.
Methods in Enzymology
|August 9, 2015
Summary
This study details a new protocol for mapping N6-methyladenosine (m(6)A) modifications across the transcriptome. The method uses immunoprecipitation and sequencing to detect m(6)A RNA, filling a critical gap in high-throughput analysis.
Area of Science:
- Molecular Biology
- Genomics
- Epigenetics
Background:
- N6-methyladenosine (m(6)A) is the most prevalent internal RNA modification.
- Accurate mapping of m(6)A modifications is crucial for understanding its regulatory roles.
- Existing methods lack the throughput, resolution, or unbiased nature required for comprehensive methylome analysis.
Purpose of the Study:
- To develop and present a detailed protocol for unbiased, high-throughput, and high-resolution m(6)A methylome mapping.
- To address the limitations of current RNA modification detection techniques.
Main Methods:
- Isolation and sequencing of m(6)A-methylated RNA fragments.
- Integration of methylated RNA immunoprecipitation (mRIP) with massively parallel sequencing.
- Utilizing a highly specific anti-m(6)A antibody for enrichment of modified RNA fragments.
Main Results:
- A robust protocol for generating m(6)A methylome maps.
- Successful enrichment of m(6)A-modified RNA fragments.
- High-resolution mapping of m(6)A modifications across the transcriptome.
Conclusions:
- The presented method provides an unbiased, high-throughput approach for m(6)A detection.
- This protocol enables comprehensive transcriptome-wide mapping of m(6)A modifications.
- The developed technique fills a significant void in RNA epigenetics research.
Related Concept Videos
03:21RNA-seq
12.6K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
12.6K
02:24Ribosome Profiling
4.3K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
4.3K
02:01pre-mRNA Processing
58.5K
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
58.5K