RNA Methylation Overview
Although the majority of nucleic acid modification research involves 5-methylcytosine (5-mC) in DNA, RNA is also extensively modified. In fact, there exist over a hundred modifications to RNA, including 5-mC, and RNA methylation is a common and naturally-occurring event in both prokaryotic and eukaryotic organisms. RNA methylation has been observed in numerous types of RNA molecules, including mRNAs, tRNAs, and non-coding RNAs, but the function of RNA methylation remains unclear.
The ability to detect and quantify 5-methylcytosine in RNA easily, efficiently, and accurately has been challenging due to the susceptibility of RNA to the pH and temperatures used for bisulfite conversion of DNA. Zymo Research now offers a solution with the EZ RNA Methylation™ Kit, which has been specifically optimized and validated for bisulfite conversion of RNA. This technique involves treating RNA with a uniquely formulated bisulfite conversion reagent, for conversion of non-methylated cytosines into uracil without compromising the quality or integrity of the RNA sample. Methylated cytosines remain unchanged during the treatment, and the methylation profile of the RNA can be determined using RT-PCR followed by DNA sequencing.