Oligo Modifications
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N6-methyladenosine (m6A)—modulating RNA localization, structure, stability, splicing, and translation

Quick facts:

Availability: RNA

Location: 5', internal, and 3′

Scales: 250 nmol to large scale

Purification: Standard desalt or HPLC

Ordering: Please email all requests for m6A to noncat@idtdna.com

A popular non-catalog modification

We welcome requests for modifications that do not appear in our website listing, and these are regularly reviewed for approval by our specialty oligo synthesis committee. One such popular modification is N6-methyladenosine (m6A). This is a modified RNA base with a methyl group at the N6 position (Figure 1).

Structure of N6-Methyl rA

Figure 1. Structure of N6-methyladenosine.


First described in the 1970s, m6A is the most prevalent internal (non-cap) mRNA modification found in higher eukaryotes [1]. It also occurs in other RNA species (tRNA, rRNA, snoRNA, lncRNA) and in a wide variety of organisms, including insects, plants, and yeast [2–5]. It is essential for cell viability and development [4–6], and has been associated with mRNA localization, structure, stability, splicing, and translation [7].

In humans, this methylation event is a natural post-transcriptional modification carried out by a complex consisting of METTL3 and METTL14 methyltransferases, and uses the consensus sequence [G/A/U][G>A]m6AC[U>A/C] [2,3]. Human and mouse transcriptome-wide analysis studies and mapping by m6A-CLIP/IP have revealed that the majority of m6A modifications are associated with regions of high evolutionary conservation and are preferentially located in the most 3' exon, suggesting the potential for 3'UTR regulation, including alternative polyadenylation [2,8].

Demethylation of m6A modified bases also occurs naturally within cells. The fat mass and obesity associated (FTO) gene encodes an enzyme that removes the methyl group from this modified base, and overexpression of FTO results in a lower level of m6A in mRNA [2]. The AlkB homolog 5, RNA demethylase (ALKBH5) gene encodes another RNA demethylase responsible for specific demethylation of m6A residues.


How to order

To order RNA oligonucleotides containing m6A modifications, email noncat@idtdna.com with your name, organization, requested scale, purification, and sequence designs. Oligos containing this modification can be ordered with HPLC purification, or with standard desalt, provided that other modifications in the sequence do not require HPLC.

References

  1. Wang X, Lu Z, et al. (2014). N6-methyladenosine-dependent regulation of messenger RNA stability. Nature, 505(7481):117–120.

  2. Meyer KD, Saletore Y, et al (2012). Comprehensive analysis of mRNA methylation reveals enrichment in 3' UTRs and near stop codons. Cell, 149(7):1635-1646.

  3. Dominissini D, Moshitch-Moshkovitz S, et al. (2012). Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq. Nature, 485(7397):201–206.

  4. Clancy MJ, Shambaugh ME, et al. (2002). Induction of sporulation in Saccharomyces cerevisiae leads to the formation of N6-methyladenosine in mRNA: a potential mechanism for the activity of the IME4 gene. Nucl Acids Res, 30(20):4509–4518.

  5. Zhong S, Li H, et al. (2008). MTA is an Arabidopsis messenger RNA adenosine methylase and interacts with a homolog of a sex-specific splicing factor. Plant Cell. 20(5):1278–1288.

  6. Hongay CF, Orr-Weaver TL. (2011). Drosophila inducer of MEiosis 4 (IME4) is required for Notch signaling during oogenesis. Proc Natl Acad Sci USA, 108(36):14855–14860.

  7. Dominissini D, Nachtergaela S, et al. (2016). The dynamic N1-methyladenosine methylome in eukaryotic messenger RNA. Nature, 530(7591):441–446.

  8. Ke S, Alemu EA, et al. (2015). A majority of m6A residues are in the last exons, allowing the potential for 3′ UTR regulation. Genes Dev, 29(19):2037–2053.

Further reading

Oligonucleotide modifications: Choosing the right mod for your needs—Review these guidelines on selecting oligonucleotide modifications to learn how they can help you in your research.

Need a non-standard modification?—Did you know that IDT offers 89 modifications that are not listed in our online catalog? A few of the more popular ones are described along with information on how to order them. IDT will consider any modification you require. Please make such inquiries to noncat@idtdna.com.

Need a library of related DNA or RNA oligo sequences?—Build variability into your oligo sequences by incorporating Mixed Bases. We offer mixes of multiple base types as well as nonstandard and modified bases.

Increase the Tm of short, AT-rich primers and probes—Add this modified base to increase the melting temperature (Tm) of primers and probes. It is especially useful when you need to work with short, A-T rich sequences.

Inverted bases—Learn how inverted bases allow you to reverse the orientation of part of your oligo sequence or add a 5'-end restricted modification to the 3'-end of your sequence.

Author: Stephanie Youtsey is the non-catalog coordinator in the Specialty & Manufacturing Services group at Integrated DNA Technologies.

© 2016 Integrated DNA Technologies. All rights reserved. Trademarks contained herein are the property of Integrated DNA Technologies, Inc. or their respective owners. For specific trademark and licensing information, see www.idtdna.com/trademarks.


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