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Single-Base tRNA Modification Sequencing
Single-Nucleotide tRNA ac4C Modification Seq(ReadC-T-seq)
RedaC-T-seq is a base-resolution, antibody-free technique for quantifying N4-acetylcytidine (ac4C) on tRNA. A reductive RedaC-T step marks acetylated cytidine and turns it into a distinguishable read-out, separating ac4C from unmodified cytidine, so each ac4C position is pinpointed at single-nucleotide resolution.
Arraystar provides a complete sample-to-data workflow for tRNA ac4C profiling, spanning RNA quality control, tRNA sample processing, library prep, sequencing, and downstream analysis. Detected ac4C sites are localized within mature tRNA transcripts and annotated by anticodon, isotype, and structural domain.
Benefits
In tRNA, the modification stabilizes the folded L-shaped structure and fine-tunes anticodon–codon decoding, so base-resolution mapping within maturation loops and the anticodon is what distinguishes it.
Single-base resolution: pinpoints ac4C sites in tRNA.
Quantitative measurement: the conversion read-out yields reliable ac4C modification fractions.
No antibody dependence: eliminates enrichment bias and cross-reactivity concerns.
Functional annotation: Sites are localized within mature tRNA transcripts with relative coordinates, tRNA gene attribution, anticodon and isotype, and structural-domain localization.
Application-ready: supports decoding fidelity, codon use, aminoacylation, and tRNA structural biology across tRNA isotypes.
| Service Name | Price |
|---|---|
| Single-Nucleotide tRNA ac4C Modification Seq(RedaC-T-seq) |
Background
In tRNA, the modification stabilizes the folded L-shaped structure and fine-tunes anticodon–codon decoding, so base-resolution mapping within maturation loops and the anticodon 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 method was validated for tRNA, sites are localized within mature tRNA transcripts with relative coordinates, tRNA gene attribution, anticodon and isotype, and structural-domain localization. (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: RedaC-T-seq pinpoints ac4C across tRNA at single-base resolution. The method uses reductive conversion to mark acetylated cytidine as a readable signature, separating it from unmodified cytidine. Because it is antibody-free, the read-out delivers quantitative modification maps of ac4C across the tRNA transcriptome.
Figure 1. RedaC-T-seq ac4C detection scheme. A reductive step specifically labels acetylated cytidine, translating it into a detectable signal that reports ac4C positions quantitatively at base resolution.
Workflow
A streamlined workflow takes tRNA samples from receipt to base-resolution ac⁴C datasets:
Figure 2. Single-Nucleotide tRNA ac4C Modification Seq workflow.
Bioinformatics
Downstream analysis maps reads to a curated tRNA reference, identifies RedaC-T conversion signatures, and reports ac4C stoichiometry per position. The pipeline also delivers motif analysis, differential modification testing, and genome-browser track files.
Deliverables
- Raw ac⁴C tRNA sequencing FASTQ data files, formatted for GEO/SRA public database submission.
- QC report with BAM alignment files and key mapping metrics.
- High-confidence single-base ac⁴C tRNA site tables (XLSX) with coordinates, anticodon, isotype, and stoichiometry.
- Single-base ac⁴C distribution plots across tRNA domains and consensus ac⁴C motif logos (PDF/PNG).
- Single-base resolution differential ac⁴C tRNA site tables (XLSX) with fold change and significance.
- Genome browser-compatible ac⁴C signal track files (bigWig/bedGraph) and a structured tRNA report.
Research Applications
- tRNA decoding fidelity: maps the modification within anticodon and wobble regions.
- Codon use & aminoacylation: relates base-resolution modification to charging and translation.
- tRNA structural biology: localizes sites within D-loop, anticodon, TΨC, and acceptor stem.
- Isotype-resolved analysis: compares modification across tRNA isotypes and isoacceptors.
- Biomarker discovery: identifies differentially modified tRNA sites in disease.
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
Sample Storage
For cells/tissue: use TRIzol or an RNA-stabilizing reagent, quick-freeze in liquid nitrogen, and keep at –80 °C.
For RNA: dissolve in ethanol or RNase-free water, store at –80 °C, and limit freeze–thaw cycles.
Dispatch Guidelines
Transfer each sample into a 1.5 mL nuclease-free tube.
Close the tube securely with parafilm or a cap lock to preserve integrity.
Send on dry ice with sufficient insulation to sustain the required temperature.
| Sample Type | Notes |
|---|---|
| Whole blood | Collect in EDTA tubes only; heparin interferes with downstream processing. |
| Cultured cells | Cell pellets are preferred to maximize material quality. |
| Tissue | Send fresh or frozen tissue; avoid necrotic regions. |
| Total RNA | Ensure OD260/280 ≥ 1.8, RIN ≥ 7, and no visible degradation; submit ≥ 10–300 µg total RNA. |
FAQ
Which tRNA features are annotated?
Sites are localized within mature tRNA transcripts with relative coordinates, tRNA gene attribution, anticodon and isotype, and structural-domain localization. This lets you interpret each site in the context of its host tRNA isotype and structural domain. The method supports both mRNA and tRNA workflows, so you can choose the readout that matches your study.
Does RedaC-T-seq measure ac4C 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 tRNA.
How does this method differ from antibody-based tRNA 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 tRNA, 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.
What research value does ac4C profiling bring?
By reporting ac4C at base resolution across tRNA, the service enables researchers to study structural and decoding roles, strengthening the evidence behind high-impact work.
What makes single-base ac4C detection important?
Accurate, quantitative ac4C signals on tRNA support dependable conclusions in translational research, offering a valued tool for RNA biology.
!This assay is provided for research applications only and is not intended for clinical diagnosis, treatment, or personal health evaluation.