RNA Methylation

What Is RNA Methylation?

RNA methylation is a reversible post-translational modification to RNA that epigenetically impacts numerous biological processes and regulates gene expression. It occurs in different RNAs, 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.

What Are Some Specific RNA Modifications?

N6-methyladenosine (m⁶A) is the most common and abundant methylation modification in RNA molecules present in eukaryotes and accounts for more than 80% of all RNA methylation. It may be referred to as the “fifth RNA base” and has broad roles in regulating embryonic development and cell fates. In particular, m6A methylation is thought to be important for messenger RNA (mRNA) regulation due to enrichment of m⁶A at 3’UTRs. It has also been linked to cancers, including leukemia, breast cancer, and prostate cancer.

N6-methyladenosine (m⁶A)

5-methylcytosine (5-mC) in RNA is a post-transcriptional epigenetic mark that is also found to be stable and common in various RNA molecules. Researchers are able to perform RNA bisulfite conversion and RT-PCR amplification, as well as cloning, and sequencing to gain information about RNA cytosine methylation states. By treating RNA with bisulfite, cytosine residues are deaminated to uracil while leaving 5-methylcytosine intact.

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. Furthermore, loss of 5-mC in vault RNAs causes aberrant processing into Argonaute-associated small RNA fragments that can function as microRNAs. Researchers continue to investigate m6A and 5-mC in RNA to uncover the mysteries of RNA methylation.

To assist you with your studies, EpigenTek offers a complete line of convenient kits to study both RNA and 5-mC methylation. Our product line includes ELISA kits to quickly measure global levels of m6A or 5mC in RNA, meRIP kits to enrich and capture m6A RNA, and ELISA-based assays to rapidly measure m6A methylase activity and m6A demethylase activity.

For additional reading on RNA methylation, check out:

• Benchtop Tip/Guide for RNA Methylation
• How CUT&RUN Helps Improve Methylated RNA Immunoprecipitation
• The Role of m6A RNA Modification in Cancer
• End-to-End Workflow for Viral and Host RNA Modification Analysis
• A Review on the Emerging Field of m6A RNA Methylation

Featured RNA Methylation Kits

	m6A RNA Methylation Quantification Kit Cat #P-9005	CUT&RUN m6A RNA Enrichment (MeRIP) Kit Cat #P-9018	5-mC RNA Methylation ELISA Easy Kit Cat #P-9009
Purpose	An ELISA-like format for absorbance-based quantitation of N6-methyladenosine in RNA	RNA immunoprecipitation (MeRIP) based on CUT&RUN for increased specificity and signals	An ELISA-like format for fluorescence-based quantitation of 5-mC RNA methylation
Key Points	Detect m6A RNA methylation in any species Colorimetric ELISA-like assay using an m6A antibody Detection limit as low as 10 pg of m6A	High Enrichment of m6A Low Sample Requirement Minimal MeRIP/sequencing background	Large signal window, low background noise Detect 5-mC RNA methylation in any species High specificity to 5-mC, with no cross-reactivity
Starting Material	Total RNA	Total RNA	Nuclear Extracts, Purified Enzymes
Lowest Input Amount	100 ng	1 µg - 20 µg	50 ng to 300 ng
Total Protocol Time	< 4 hours	< 3 hours	< 3 hours
Reactions	48,96	24	8,96
Vessel Format	96-Well Plate	Columns/Tubes	96-Well Plate