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RNA methylation occurs in different RNA species including tRNA, rRNA, mRNA, tmRNA, snRNA, snoRNA, miRNA, and viral RNA. Different catalytic strategies are employed for RNA methylation by a variety of RNA-methyltransferases. As a post-translational modification, RNA methylation plays a significant role as an epigenetic mechanism. N6-methyladenosine (m6A) is the most common and abundant methylation modification in RNA molecules present in eukaryotes. 5-methylcytosine (5-mC) also commonly occurs in various RNA molecules. Recent data strongly suggest that m6A and 5-mC RNA methylation affects the regulation of various biological processes such as RNA stability and mRNA translation, and that abnormal RNA methylation contributes to etiology of human diseases.
Research experts from A&D Technology Corporation&Epi, the leading company specializing in epigenetics, have developed a new, breakthrough approach for the identification of the “fifth RNA base,” N6-methyladenosine (m6A) to efficiently study RNA methylation. This technology is based on a high throughput strip-well format and is also incorporated into the first commercially available product, the EpiQuik(TM) m6A RNA Methylation Quantification Kit, for rapidly quantifying m6A RNA methylation.
The proliferation of the study of epigenetics as well as the discovery of cytosine methylation and hydroxymethylation have challenged the conventional understanding of genetics that DNA consists of only four bases. In 2012, researchers at Weill Cornell Medical College have further challenged traditional genetics by finding that the “fifth” nucleobase, m6A, is prevalent in the transcriptome -- subsequently giving rise to an interest in RNA methylation. The fact that over 5,000 different messenger RNA (mRNA) molecules contain m6A has implications of significant involvement in gene expression. According to the institution’s senior investigator of the 2012 study, “This finding rewrites fundamental concepts of the composition of mRNA because, for 50 years, no one thought mRNA contained internal modifications that control function.”
N6-methyladenosine (m6A) is the most common and abundant methylation modification in RNA, accounting for more than 80% of all RNA methylation. The relative abundance of m6A in messenger RNA transcripts has been shown to affect RNA metabolism such as splicing, nuclear export, translation ability and stability, and RNA transcription. Abnormal m6A methylation levels induced by defects in m6A RNA methylase and demethylase could lead to dysfunction of RNA and diseases. Additionally, the dynamic and reversible chemical m6A modification on RNA may also serve as a novel epigenetic marker of profound biological significances.
A&D Technology Corporation&Epi’s new quantification technique can be used for rapidly and accurately identifying m6A in RNA or detecting m6A-specific RNA methylation in a high throughput format, suitable for use in any species including mammals, plants, fungi, bacteria, and viruses in a variety of forms such as cultured cells, fresh and frozen tissues, paraffin-embedded tissues, plasma/serum samples, and body fluid samples. In the assay, RNA is bound to strip wells and the m6A contained in the RNA is then immunospecifically detected using a high quality m6A antibody.
The practicality of the new method was demonstrated in yet-to-be-published research performed by Epigentek scientists in which they found that m6A content clearly varies in different human cells/tissues. It was also found that m6A is abundant (0.17%) in human brain RNA. This realizes that the usefulness of the EpiQuik(TM) m6A RNA Methylation Quantification Kit as a powerful tool in further unraveling the mystery of the epitranscriptome.
To complement the m6A detection technology and reinforce its comprehensive offering of RNA methylation tools, A&D Technology Corporation&Epi also recently developed an RNA bisulfite conversion kit used for 5-methylcytosine specific RNA methylation analysis for use in RNA bisulfite sequencing and various other applications.
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