RiboMethSeq

RiboMethseq is a sequencing-based technology used to map RNA modifications, specifically methylation, within RNA molecules. It is an adaptation of the more general RNA modification sequencing (Rimseq) approach. The core principle of RiboMethseq involves treating RNA with a specific chemical reagent, typically N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide methiodide (CMS), which reacts with methylated bases. This reaction introduces a specific chemical modification to the RNA at the methylated position.

Following the chemical treatment, the RNA is fragmented and then subjected to standard next-generation sequencing protocols. During the sequencing process, the modified bases due to CMS treatment are detected as specific sequencing errors or deviations from the expected base calls. For example, a methylated adenosine might be read as a guanosine due to the chemical modification. By analyzing these characteristic "errors" across the sequenced reads, researchers can precisely pinpoint the locations of methylated bases within the RNA transcript.

This method is valuable for studying the epitranscriptome, the collection of all RNA modifications in a cell. Understanding RNA methylation is crucial as these modifications can influence various aspects of RNA biology, including stability, translation efficiency, and splicing. RiboMethseq allows for high-throughput and genome-wide mapping of these modifications, providing insights into their roles in cellular processes and disease. Its sensitivity and specificity enable the detection of even low-abundance modifications.