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Single-Base RNA Modification Sequencing
Single-Nucleotide RNA ac4C Modification Seq(ReadC-T-seq)
Single-Nucleotide RNA ac4C Modification Seq(RedaC-T-seq) is an antibody-free, base-resolution method. A reductive RedaC-T conversion selectively acts on acetylated cytidine and records it as a readable signature while distinguishing unmodified cytidine, mapping each ac4C site at single-base resolution. This maps and quantifies ac4C sites across protein-coding transcripts at single-base resolution.
Arraystar Single-Nucleotide RNA ac4C Modification Seq(RedaC-T-seq) is an end-to-end sample-to-data service for mRNA, from RNA sample QC and mRNA treatment through library construction, sequencing, and bioinformatics. Sites are annotated to the reference transcriptome with genomic coordinates, host gene, and functional region attribution.
Benefits
In mRNA, the modification shapes transcript stability, translation, and gene regulation, so base-resolution mapping across coding and untranslated regions is what distinguishes it.
Single-base resolution: pinpoints ac4C sites in mRNA.
Quantitative stoichiometry: conversion-based readouts give accurate modification ratios.
Antibody-free and unbiased: avoids enrichment cross-reactivity and pull-down bias.
Functional annotation: Sites are annotated to the reference transcriptome with genomic coordinates, host gene, and functional region attribution.
Application-ready: supports translation control, RNA stability, coding-region function, and gene-regulation studies across the mRNA transcriptome.
| Service Name | RNA Class | Price |
|---|---|---|
| Single-Nucleotide RNA ac4C Modification Seq(RedaC-T-seq) | mRNA | |
| Single-Nucleotide RNA ac4C Modification Seq(RedaC-T-seq) | mRNA & lncRNA |
Background
In mRNA, the modification shapes transcript stability, translation, and gene regulation, so base-resolution mapping across coding and untranslated regions is what distinguishes it. A reductive RedaC-T conversion selectively acts on acetylated cytidine and records it as a readable signature while distinguishing unmodified cytidine, mapping each ac4C site at single-base resolution. This antibody-free readout maps each ac4C site at single-base resolution. The same workflow extends to long non-coding RNAs (lncRNAs), enabling researchers to profile this modification on lncRNA transcripts in parallel with mRNA in a single assay.
The method was validated for mRNA; sites are annotated to the reference transcriptome with genomic coordinates, host gene, and functional region attribution. (Arango D, Sturgill D, Yang R, et al. Oberdoerffer S. Molecular Cell 2022;82(15):2797-2814.e11., DOI: 10.1016/j.molcel.2022.05.016).
Key concept: Single-Nucleotide RNA ac4C Modification Seq(RedaC-T-seq) is a base-resolution sequencing method in which a reductive redac-t conversion selectively acts on acetylated cytidine and records it as a readable signature while distinguishing unmodified cytidine, mapping each ac4c site at single-base resolution. This antibody-free readout yields quantitative, single-base modification maps across protein-coding transcripts.
Figure 1. RedaC-T-seq ac4C detection scheme. Reductive conversion selectively acts on acetylated cytidine and records it as a readable signature, mapping and quantifying ac4C at single-base resolution.
Workflow
Five steps from RNA sample to single-base N4-acetylcytidine (ac⁴C) data:
Figure 2. Single-Nucleotide RNA ac4C Modification Seq workflow.
Bioinformatics
The bioinformatics pipeline aligns reads to the reference transcriptome, recognizes RedaC-T conversion signatures, and quantifies ac4C stoichiometry at single-nucleotide resolution. Analyses include motif characterization, differential modification, functional enrichment, and genome-browser visualization.
Deliverables
- Raw ac⁴C mRNA sequencing FASTQ data files, formatted for GEO/SRA public database submission.
- Sample and sequencing QC reports with sorted BAM alignment files and core mapping statistics.
- High-confidence single-base ac⁴C site annotation tables (XLSX) with coordinates, genes, and absolute stoichiometry.
- Single-base ac⁴C transcript feature distribution figures and consensus ac⁴C motif logos (PDF/PNG format).
- Single-base resolution differential ac⁴C methylation tables (XLSX) with fold change and significance metrics.
- Gene Ontology (GO) enrichment analysis reports for ac⁴C-modified genes, provided in HTML/PDF/PNG formats.
- Genome browser-compatible track files (bigWig/bedGraph) and a full structured ac⁴C analysis project report.
Research Applications
- Transcriptome-wide mapping of the modification across protein-coding and non-coding mRNA.
- Translational control: relates base-resolution modification to mRNA translation and stability.
- Coding-region function: localizes sites within CDS, UTR, and splice-adjacent regions.
- Disease-oriented studies: finds differentially modified mRNA sites as biomarkers.
References
[1] Arango D, Sturgill D, Yang R, et al. Oberdoerffer S. Direct epitranscriptomic regulation of mammalian translation initiation through N4-acetylcytidine. Molecular Cell 2022;82(15):2797-2814.e11. DOI: 10.1016/j.molcel.2022.05.016.
Sample Requirements
Storage Guidelines
Cells and Tissues: Preserve in TRIzol or an RNA stabilization solution; snap freeze in liquid nitrogen and store at –80 °C.
RNA: Resuspend in ethanol or RNase-free ultrapure water; store at –80 °C and avoid multiple freeze-thaw cycles.
Shipping Instructions
Place the sample in a 1.5 mL RNase-free microcentrifuge tube.
Seal the tube with parafilm or a cap lock to ensure sample integrity.
Ship the package on dry ice with adequate insulation to maintain the required temperature.
| Sample Type | Notes |
|---|---|
| Whole blood | Use EDTA tubes only, as heparin is not compatible with subsequent analytical procedures. |
| Cultured cells | Submission of cell pellets is preferred to ensure high-quality material for processing. |
| Tissue | Provide fresh or frozen specimens, ensuring that necrotic material is strictly avoided. |
| Total RNA | Maintain an OD 260/280 ratio ≥ 1.8 and RIN ≥ 7 with no visible degradation. Submit at least 10–300 µg total RNA. |
FAQ
Which mRNA features are annotated?
Sites are annotated to the reference transcriptome with genomic coordinates, host gene, and functional region attribution. This lets you interpret each site in the context of its host gene and mRNA functional region. The method supports both mRNA and tRNA workflows, so you can choose the readout that matches your study.
Can this method quantify the modification at single-base resolution?
Yes. A reductive RedaC-T conversion selectively acts on acetylated cytidine and records it as a readable signature while distinguishing unmodified cytidine, mapping each ac4C site at single-base resolution. This provides single-base resolution and absolute stoichiometry for each site in mRNA.
How does this method differ from antibody-based mRNA profiling?
Antibody-based methods rely on enrichment and offer limited resolution with intrinsic motif bias. This method is antibody-free and delivers base-resolution, quantitative calling in mRNA, avoiding cross-reactivity and enrichment bias. This suits projects needing quantitative site-level mapping rather than region-level enrichment, and it pairs well with orthogonal validation.
How does this service support research on ac4C biology?
By resolving ac4C at single-base resolution, this service lets investigators examine the regulatory roles of ac4C in gene expression, translation, and disease, providing a strong evidence base for high-impact mechanistic studies.
Why is high-resolution ac4C profiling a valuable research asset?
Precise, quantitative ac4C mapping underpins reliable conclusions across translational fields, making it a trusted tool for advancing understanding of RNA regulation and its biological consequences.
Does this service also profile lncRNA?
Yes. We provide two service options: mRNA modification, and mRNA plus long non-coding RNA (lncRNA) modification. Both transcript types can be analyzed together in a single experiment, so there is no need to split them into two projects, and the scope of your study opens up accordingly.
!This assay is provided for research applications only and is not intended for clinical diagnosis, treatment, or personal health evaluation.