What is 2'-O-Methylation

Methylation of the ribose 2'-hydroxyl group (2'-O-Me), is among the most common RNA covalent modifications, 2'-O-methylated (Nm) is abundant in rRNA, tRNA, snRNA and microRNA; and it is essential for the biogenesis, metabolism and function of these molecules. It is catalyzed by an ensemble of specialized enzymes known as RNA methyltransferases, their delicate choreography governed by the transfer of a methyl group from the esteemed methyl donor molecule, S-adenosyl methionine (SAM), to the target ribose residue.

Importance of 2'-O-Methylation

RNA Stability and Structure: The Guardians of Molecular Integrity

As the guardians of molecular integrity, 2'-O-methyl groups elevate RNA to the pinnacle of stability. Their strategic presence fends off the ravages of nucleases, acting as molecular sentinels against degradation. A tapestry of protection is woven, safeguarding RNA's delicate threads from the ceaseless onslaught of molecular adversaries. But the story doesn't end there. These modifications don the mantle of structural influencers, intricately shaping RNA's fold and tertiary conformation. Interactions, both subtle and profound, unfurl as the dance of 2'-O-methylation intertwines with the choreography of proteins and small RNAs, molding RNA's structural destiny.

RNA Processing and Splicing: Guiding the Symphony of RNA Maturation

A symphony of RNA maturation unfolds, guided by the deft hand of 2'-O-methylation. Within the intricate realms of snRNAs and snoRNAs, these modifications act as signposts, directing the recognition and procession of pre-mRNA. The orchestra of RNA processing and splicing harmonizes flawlessly, ensuring the precise stitching of exons and the generation of mature RNA transcripts. A meticulous choreography, choreographed by the masterful touch of 2'-O-methylation, brings forth a symphony of gene expression, seamlessly translating the blueprint of life.

Translation Regulation: The Language of Protein Synthesis Translated

In the realm of translation, where RNA metamorphoses into the language of proteins, 2'-O-methylation emerges as a silent conductor, silently influencing the symphony. At specific sites within the mRNA molecule, these modifications orchestrate an intricate ballet, dictating the efficiency of translation and the fidelity of protein synthesis. The dance of 2'-O-methylation whispers subtle cues to the ribosome, shaping the destiny of protein synthesis and sculpting the intricate web of cellular function.

Immune Response Modulation: Unveiling the Viral Strategies

In the realm of the immune response, a captivating tale unfolds—one where 2'-O-methylation becomes a key player in the delicate balance between host and pathogen. Recent investigations illuminate the stealthy tactics employed by certain RNA viruses, harnessing the power of 2'-O-methylation to evade detection and evade the watchful eyes of the immune system. This molecular cloak empowers these viral interlopers, enabling their replication and propagation within unsuspecting host cells. The profound understanding of these intricate mechanisms unfurls avenues for novel antiviral strategies, a shield forged from the secrets of 2'-O-methylation.

Nm-seq for 2'-O-Methylation Detection

Accurate detection and characterization of 2'-O-methylation are essential for studying its functional roles and identifying disease-associated alterations. In recent years, a novel technique called Nm-seq has emerged as a powerful tool for detecting 2'-O-methylation sites in RNA molecules. Despite advances in high-throughput detection, the inert chemical nature of Nm still limits sensitivity.

CD Genomics provides Nm-seq, which leverages oxidative cleavage of ribose 2',3'-vicinal diols by periodate to expose, enrich and map Nm sites in the transcriptome without bias and with single-nucleotide precision.

Figure 1. A schematic illustration of Nm-seq (Dai, 2017)

Specific Nm-seq Approaches We Offer

Methods

• First internal Nm sites in RNA fragments were exposed by iterative oxidation–elimination–dephosphorylation (OED) cycles
• A final round of oxidation–elimination (OE) reaction, without dephosphorylation, is then performed to generate two types of 3' ends that differ in their ligation compatibility
• Fragments ending with Nm are preferentially ligated to the 3' adaptor
• Enriched by PCR amplification and further sequencing

Our Nm-seq are available for a wide range of sequencing applications:

• Complete transcriptome sequencing of Nm
• Nm LnRNA sequencing
• Nm mRNA sequencing

Our bioinformatics analysis services include, but are not limited to:

• Sequencing data quality control
• Comparison with reference genome
• Identified methylated sites
• Length distribution of methylated sites and motif analysis results
• Differential methylated sites
• GO and Pathway analysis results of differential methylated sites

Service Advantages

• Single base localization of Nm
• Nm detection in the whole transcriptome
• Targeted detection of Nm of different types of RNA molecules

Our Capabilities

• Wide detection range: Whole-transcriptome wide Nm detection.
• Flexible sample requirements: Cells, tissues or RNA are acceptable
• Specialized bioinformatics analysis: Strong bioinformatic team, professional Nm data analysis.

Sample Requirements

• Cell samples

The number of cells shall not be less than 1×10⁸.

• Fresh tissue samples

The quantity of the tissue samples should be between 500 mg - 5g.

• RNA samples

The quantity of RNA sample should be no less than 100 μg, RNA purity: OD260/280: 1.6-2.3, no obvious degradation of RNA.

Service Process

Deliverables

• Related experimental results raw data
• Experimental report
• Data analysis
• Image and result analysis
• Bioinformatics analysis results
• Details in 2'-O-methylation sequencing for your writing

Our Features

Why Choose Us?

CD Genomics is a company that provides professional and comprehensive Nm-seq services. We can conduct high-throughput detection and single base localization of 2 '- O-RNA modification. CD Genomics can provide you with personalized solutions to help you thrive every step of the way around your interest in your workflow. If you would like to know more about this service, please feel free to contact us.

Reference

1. Dai, Qing, et al. Nm-seq maps 2'-O-methylation sites in human mRNA with base precision. Nature methods 14.7 (2017): 695-698.