Related Experiment Video
Updated: Feb 10, 2026

Characterizing RNA Modifications in Single Neurons Using Mass Spectrometry
Published on: April 21, 2022
Probing N⁶-methyladenosine (m⁶A) RNA Modification in Total RNA with SCARLET
Abstract:
Posttranscriptional N (6)-methyladenosine (m(6)A) RNA modification is indispensable for cell development and viability; however, functional investigation of m(6)A biological function has been hindered by the lack of methods for its precise identification and quantitation. Here, we describe a method that accurately identifies m(6)A position and modification fraction in human messenger RNA (mRNA) and long noncoding RNA (lncRNA) at single-nucleotide resolution, termed as "site-specific cleavage and radioactive-labeling followed by ligation-assisted extraction and thin-layer chromatography (SCARLET)" (Fig. 1). This method combines two previously established techniques, site-specific cleavage and splint ligation, to probe the m(6)A RNA modification status at any mRNA/lncRNA site in the total RNA pool.
Related Concept Videos
Ribosomal RNA Synthesis
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
RNA Interference
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
RNA Structure
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...
RNA Stability
Transfer RNA Synthesis
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
RNA Splicing

