Description
Key facts
Detection method:Colorimetric,
Sample types:DNA,
Assay type:Quantitative,
Assay Platform:Microplate reader
Product details:
m6A DNA Methylation Assay Kit (Colorimetric)(ab233488) kit contains all reagents necessary for the quantification of m6A in DNA. In this assay, DNA is bound to strip wells using DNA high binding solution. m6A is detected using capture and detection antibodies. The detected signal is enhanced and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer. The amount of m6A is proportional to the OD intensity measured.
N 6 -methyladenosine (m6A) is the most common and abundant modification on RNA molecules present in eukaryotes. DNA m 6A is also identified in multicellular eukaryotes including Caenorhabditis elegans and Drosophila melanogaster, and furthermore identified in higher eukaryotes including plants, mouse and human cells. m6A plays crucial roles in regulating DNA replication, transposition, transcription, and cellular defense. In humans, the DNA m 6A modification is most likely catalyzed by a methyltransferase complex METTL3 and removed by the α-ketoglutarate (α-KG)- and Fe2+ -dependent dioxygenases such as ALKBH5 and TET-like enzymes. It was shown that METTL3 and α-KG /Fe2+ - dependent dioxygenases play important roles in many biological processes, ranging from development and metabolism to fertility.
The dynamic and reversible chemical m6A modification on DNA may also serve as a novel epigenetic marker of profound biological significance. Down-regulation of m 6A modification was first characterized in human cancer cells and tissues, relative to their normal controls. m 6A is found to be the most regulated DNA modification in cancers. In addition to the regulation in cancer cells, relative to the primary cell/tissues which contain quite low amounts of DNA m 6A (<0.001%), a hundreds-fold increase of m 6A modification was found for in vitro cultured human cells (0.03%-0.22%). Therefore, identifying m 6A DNA methylation levels and distribution on DNA could advance understanding of epigenetic regulation of biological process at the genomic level, and further provide useful information for improving diagnostics and therapeutics of disease.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
M6A DNA methylation also known as N6-methyladenine modification involves the addition of a methyl group at the nitrogen-6 position of adenine within DNA strands. This modification mainly occurs in the eukaryotic genome and plays roles in gene expression regulation. The exact mass of m6A itself cannot be specified as it is a chemical modification rather than a separate protein. m6A methylation events are observed in various organisms including humans mice and plants suggesting its evolutionary significance across different species.
Biological function summary
The m6A modification of DNA affects transcriptional activity by influencing the interaction between DNA and proteins. This type of methylation acts similarly to a switch turning gene expression on or off as needed. In several cases m6A works closely within complexes involving RNA-binding proteins and transcription factors coordinating the stability splicing and translation of mRNAs. This suggests a significant role in post-transcriptional gene regulation. For instance components such as METTL3 and METTL14 are associated with m6A methylation and contribute to forming a complex that performs these regulatory functions.
Pathways
The modification by m6A DNA methylation appears to be an integral part of epigenetic pathways that govern cellular differentiation and development. It also influences the methylation signaling pathway aiding in the fine-tuning of epigenetic states within the cell. Proteins related to m6A-modified pathways include ALKBH5 and FTO both of which demethylate m6A marks allowing dynamic control of DNA interpretation. Understanding these modifications within pathways requires considering how they affect gene regulatory networks on an epigenetic level.
Altered m6A DNA methylation patterns have been linked to various cancers and neurological disorders. Dysregulation of m6A has been observed in cancers such as acute myeloid leukemia where it is implicated in the proliferation and differentiation of malignant cells. The proteins METTL3 and FTO play roles in cancer progression making them targets for therapeutic intervention. In neurological disorders defective m6A methylation has associations with autism spectrum disorder where it affects neural development and brain function. The modification's regulatory impact on neurodevelopmental genes highlights its potential involvement in these conditions.
