RNA Internal N7-Methylguanosine (m7G) Analysis

Internal N7-methylguanosine (m7G) is a pivotal yet challenging-to-study RNA modification that plays crucial roles in transcript processing, stability, and translation. CD BioSciences provides specialized, high-resolution m7G analysis services, leveraging orthogonal technologies to overcome detection hurdles and deliver precise mapping and quantification of this important epitranscriptomic mark.

Introduction to Internal N7-Methylguanosine (m7G)

N7-methylguanosine (m7G) is a positively charged methylation occurring at the nitrogen-7 position of guanine. While famously forming the 5' cap structure of eukaryotic mRNA, it also exists as a widespread internal modification within transfer RNA (tRNA), ribosomal RNA (rRNA), and messenger RNA (mRNA), where its detection has been historically difficult due to chemical instability and lack of specific antibodies. Functionally, internal m7G is essential for maintaining tRNA structural integrity and accurate decoding, influences rRNA biogenesis and function, and is increasingly recognized as a regulator of mRNA metabolism, potentially affecting splicing, export, and translation efficiency. Its dysregulation is linked to human diseases, including cancers and neurodevelopmental disorders, making precise mapping and quantification vital for uncovering novel regulatory pathways and therapeutic targets in the epitranscriptome.

Fig.1 The cellular m7G modification machinery. (Luo Y, et al., 2022)

Our Services

Building on a deep understanding of epitranscriptomic complexity, CD BioSciences addresses the specific analytical challenges of internal m7G by offering an integrated service platform. We combine robust chemical biology techniques with state-of-the-art mass spectrometry to provide unambiguous detection, precise quantification, and transcriptome-wide mapping, empowering researchers to confidently explore the functional landscape of this elusive modification.

Comprehensive m7G Analysis Solution

1. Borohydride Reduction Sequencing (BoRed-seq): Utilizes specific sodium borohydride reduction to convert internal m7G into a stable, sequence-able lesion, enabling its transcriptome-wide mapping at single-nucleotide resolution without antibodies.
2. m7G Methylated RNA Immunoprecipitation Sequencing (m7G-MeRIP-seq): Employs optimized protocols with validated antibodies for the immunoprecipitation and sequencing of m7G-modified RNA fragments.
3. Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS): Provides absolute, quantitative analysis of m7G nucleosides, distinguishing internal m7G from the abundant 5' cap m7G through specific enzymatic digestion or fragmentation strategies.

Workflow of RNA m7G Analysis Service

Sample & Application Support

Supported Sample Types

Our optimized protocols are compatible with a wide range of high-quality RNA inputs to accommodate diverse research needs. We routinely analyze samples including:

1. Cells & Tissues: Cultured cell lines, primary cells, animal tissues, plant tissues, frozen tissue specimens.
2. Clinical & Pathological Samples: Whole blood, PBMCs, cell-free plasma/serum, FFPE samples.
3. Specific RNA Fractions: Poly(A)+ mRNA, tRNA, rRNA, other non-coding RNA subsets.
4. Synthetic RNA: In vitro transcribed (IVT) RNA, other engineered RNA.

Key Application Areas

The RNA m7G analysis services are designed to empower research and development across several critical fields:

1. Fundamental Mechanism Discovery: Uncover the role of internal m7G in regulating RNA processing, stability, translation, and its interplay with other epigenetic marks.
2. Disease Biomarker Identification: Investigate the differential methylation landscape of m7G in diseases such as cancer, neurological disorders, and metabolic conditions to discover novel diagnostic or prognostic markers.
3. Therapeutic Target & Drug Development: Profile m7G modifications to identify new therapeutic targets and assess drug efficacy on the epitranscriptome.
4. Synthetic Biology & mRNA Therapeutics Optimization: Characterize and optimize m7G deposition in engineered or therapeutic RNA molecules to modulate their stability, immunogenicity, and translational efficiency.

Our Advantages

1. Guarantee Specificity & Accuracy: We overcome the technical challenge of distinguishing internal m7G from the 5' cap m7G by employing orthogonal, validated methods like BoRed-seq and LC-MS/MS, ensuring high-confidence results.
2. Offer Unmatched Technical Expertise: Our team provides end-to-end guidance, from selecting the optimal protocol for your sample type to interpreting complex data, leveraging deep specialization in challenging RNA modifications.
3. Ensure Robustness Across Sample Types: Our rigorously optimized workflows deliver reliable performance for a wide range of inputs, from common cell lines to challenging clinical specimens like FFPE and cell-free RNA.
4. Drive Actionable Biological Insights: We go beyond data delivery by providing integrated analysis and clear biological interpretation, linking m7G dynamics to functional outcomes in gene regulation and disease.

CD BioSciences provides a comprehensive portfolio of epitranscriptomic analysis services beyond internal m7G profiling. Our expertise also encompasses m6A, m5C, ac4C, Ψ, m6Am and m1A analysis, as well as RNA adenosine-to-inosine (A-to-I) editing analysis. We are committed to providing fully customized end-to-end solutions, whether your goal is basic research, biomarker discovery, or therapeutic drug development, we can support your specific objectives with accurate and reliable data. Please contact us for a detailed consultation.

Reference

1. Luo Y, Yao Y, Wu P, et al. The potential role of N 7-methylguanosine (m7G) in cancer[J]. Journal of hematology & oncology, 2022, 15(1): 63.