Until now, over 100 types of cellular RNA modifications have been identified in both coding and a variety of non-coding RNAs. N6-methyladenosine (m6A) is the most prevalent and abundant mRNA modification on eukaryote mRNA. The presence of m6A mRNA modifications contributes to diverse fundamental cellular functions, such as pre-mRNA splicing, nuclear transport, stability, translation, and microRNA biogenesis, implying an association with numerous human diseases.
m6A was discovered in the 1970s in a wide range of cellular mRNAs. m6A modifications are being mapped at the nucleotide level. Methylation occurs at the sixth position of nitrogen atoms of adenosine at the post-transcriptional level with S-adenosylmethionine serving as the methyl donor for m6A formation, which is termed m6A modification. m6A modifications are tied to most aspects of the mRNA life cycle. Under regulatory control by the Mettl3/Mettl14 methyltransferase complex, m6A is the most prevalent internal mRNA modification. m6A depletion by Mettl14 knockout in embryonic mouse brains prolongs the cell cycle of radial glia cells and extends cortical neurogenesis into postnatal stages. m6A depletion by Mettl3 knockdown also leads to a prolonged cell cycle and maintenance of radial glia cells.
Fig.1 Roles of the N6-methyladenosine Writer, Eraser and Reader complexes in regulating mRNA. RNA m6A modifications serve important roles in the molecular mechanisms of gene biology.1
The biological functions of RNA m6A modifications occur at three different levels: 1) At the molecular level, RNA modification posttranscriptionally regulates RNA splicing, transport, translation, stability, and localization. 2) At the cellular level, RNA m6A modifications determine the fate of mammalian embryonic stem cells (mESCs). All the findings suggest that m6A modification plays a powerful and precise regulatory role in cell developmental programs. 3) At the physiological level, reversible m6A modification has various consequences. Studies have found that m6A modification is related to neural stem cells, brain development, Parkinson's disease, and mental illness. Furthermore, m6A modification has been found to have an impact on tumor initiation and progression through various mechanisms.
The subsequent focus on researching m6A modification in the setting of physiological and pathological processes will enrich our knowledge concerning a variety of conditions, contribute to the advancement of the biological sciences and provide us with novel therapeutic strategies. With Ph.D. level scientists and extensive experience in custom mRNA synthesis and modification, we are dedicated to offering high-quality m6A mRNA modification service to meet our clients' R&D timeline and budget.
Moreover, we provide a range of methods for the analysis of m6A modifications:
A: Creative Biolabs offers a comprehensive range of methods for the analysis of m6A modifications, including semi-quantitative approaches (blot technology, methyl sensitivity of MazF RNA endonucleases, immune-northern blot), quantitative approaches (RNA photo-crosslinkers and quantification, proteomics, electrochemical immunosensor method, support vector machine-based method), and methods to detect precise locations (high-resolution melting analysis, level and isoform characterization, SCARLET, MeRIP-seq, MiClip, DNA polymerase for direct m6A sequencing, HPLC).
A: Creative Biolabs ensures high-quality results by employing Ph.D.-level scientists with extensive experience in custom mRNA synthesis and modification, alongside a meticulous approach to meet clients' R&D timelines and budgets.
A: The study of m6A modification is vital as it enhances our understanding of various physiological and pathological conditions, contributes to the advancement of biological sciences, and helps in developing novel therapeutic strategies.
A: Applications that can benefit include research on gene expression regulation, RNA stability, disease mechanism studies, therapeutic target identification, and the development of RNA-based therapies.
A: MiClip allows for individual-nucleotide-resolution cross-linking and immunoprecipitation, providing high precision in locating m6A modifications, while HPLC offers high-performance liquid chromatography for accurate and efficient separation, identification, and quantification of m6A-modified nucleotides.
A: Creative Biolabs provides a comprehensive range of m6A mRNA modification services, including methylation site mapping, m6A-RNA binding protein identification, m6A-impact functional studies, quantitative m6A profiling, custom m6A RNA immunoprecipitation (MeRIP), and high-throughput sequencing. These services help researchers elucidate the roles and mechanisms of m6A modifications in various biological contexts.
A: Creative Biolabs uses advanced techniques such as m6A RNA immunoprecipitation followed by sequencing (MeRIP-seq), mass spectrometry, and site-specific labeling to accurately detect and map m6A modifications on mRNA. These methods provide high-resolution and high-throughput data on m6A distribution and dynamics.
This study explores the role of N6-methyladenosine (m6A) mRNA modification in maintaining colonic epithelial cell homeostasis, critical for the intestinal barrier and proper function. Researchers discovered that m6A modification is essential for the homeostatic self-renewal of colonic stem cells. Deleting the methyltransferase gene Mettl14 in mouse colons led to stem cell apoptosis, mucosal barrier dysfunction, and severe colitis. Mechanistically, m6A modification, mediated by Mettl14, stabilizes Nfkbia mRNA, thereby modulating the NF-κB pathway to prevent epithelial cell death. This study reveals m6A's crucial role in colonic epithelial cells and suggests it as a potential therapeutic target for inflammatory bowel disease (IBD).
Fig.2 m6A modification regulates colonic epithelial cell homeostasis by restricting TNF-induced cell death.2
| Cat. No | Product Name | Promoter |
|---|---|---|
| CAT#: GTVCR-WQ001MR | IVTScrip™ pT7-mRNA-EGFP Vector | T7 |
| CAT#: GTVCR-WQ002MR | IVTScrip™ pT7-VEE-mRNA-EGFP Vector | T7 |
| CAT#: GTVCR-WQ003MR | IVTScrip™ pT7-VEE-mRNA-FLuc Vector | T7 |
| CAT#: GTVCR-WQ87MR | IVTScrip™ pT7-VEE-mRNA-Anti-SELP, 42-89-glycoprotein Vector | T7 |
| Cat. No | Product Name | Type |
|---|---|---|
| CAT#: GTTS-WQ001MR) | IVTScrip™ mRNA-EGFP (Cap 1, 30 nt-poly(A)) | Reporter Gene |
| CAT#: GTTS-WK18036MR | IVTScrip™ mRNA-Human AIMP2, (Cap 1, Pseudo-UTP, 120 nt-poly(A)) | Enzyme mRNA |
| (CAT#: GTTS-WQ004MR) | IVTScrip™ mRNA-Fluc (Cap 1, 30 nt-poly(A)) | Reporter Gene |
| (CAT#: GTTS-WQ009MR) | IVTScrip™ mRNA-β gal (Cap 1, 30 nt-poly(A)) | Reporter Gene |
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