tRNA Charging Seq (modification-induced misincorporation tRNA-seq) is a novel and powerful tRNA-seq that can profile tRNA expression, tRNA modifications, and tRNA charging [12]. The method uses a thermostable reverse transcriptase to synthesize full-length cDNA for tRNAs at high efficiency, allowing quantitative analysis of tRNA expression profiles.
One assay for tRNA expression, modification, and charging
As tRNA modifications induce base misincorporations during reverse transcription, tRNA Charging Seq detects the modifications at precise base positions based on the induced mutations in the tRNA sequences. By analyzing 3′-CCA end oxidation/β-elimination, the amino acid charged or uncharged status can be measured.
tRNA Charging Seq overcomes many difficulties of reverse transcription blocks in cDNA synthesis by tRNA modifications and low full-length cDNA yields with old tRNA-seq techniques. It provides tRNA abundance, tRNA modification, and tRNA charging profiles key to tRNA studies in, for example, cancer drug resistance, cardiac fibrosis, and many other diseases.
tRNA Charging Seq (modification-induced misincorporation tRNA-seq) is a novel tRNA-seq that profiles tRNA expression, tRNA modifications, and tRNA charging simultaneously. A thermostable reverse transcriptase synthesizes full-length tRNA cDNA at high efficiency; modifications induce base misincorporations that mark their positions, and 3′-CCA end analysis measures amino acid charging.
Standard tRNA Charging Seq project — tRNA expression, tRNA modification, and tRNA charging in one assay
| Service Name | Description | Price |
|---|---|---|
| tRNA Charging Seq | tRNA expression, tRNA modification, tRNA charging |
Key advantages of Arraystar tRNA Charging Seq
tRNA expression, tRNA modification, and tRNA charging are profiled in a single assay.
Highly efficient full-length cDNA synthesis to reduce mapping/counting inaccuracy.
tRNA modifications, e.g. m1A, m1G, m3C, acp3U, are predicted at single-nucleotide resolution.
Correlate tRNA charging with translation activities in the same study.
A wealth of tRNA multi-omics data with common analyses (e.g. differential analyses) and detailed annotations.
Publication-ready graphics and visualization included in every project.
Why tRNA expression, modification, and charging matter
tRNAs are abundant small RNAs that carry amino acids and decode genetic codons for protein translation in the cells [1]. In addition to the canonical protein translation function, tRNAs have been discovered more recently as active regulators in mRNA translation and diseases by various mechanisms [2-4]. tRNA expression, tRNA modification, and tRNA charging are key profiles in regulating tRNA molecular functions (Fig. 1).
tRNA expression levels profoundly impact mRNA translation. During pathogenesis, changed tRNA repertoires by differential tRNA expression directly regulate the translation efficiency and accuracy of target mRNAs by codon preferences [2]. Cell proliferation, differentiation, and apoptosis are often dynamically regulated by tRNA expression levels.
tRNA modifications exert great influence on protein translation. Certain modifications (e.g. m7G, m1A) can increase the affinity between the tRNA and ribosome to elevate mRNA translation speed and stability, whereas aberrant tRNA modifications can cause ribosome stalling or decoding errors leading to protein synthesis problems [3, 5].
tRNA charging is correlated with mRNA translation efficiency. As only the amino acid charged tRNAs can enter the ribosome and pair with the anticodons, their aminoacylation levels directly determine tRNA translation activities. When tRNA charging is low, translation efficiency is reduced, even enough to touch off mRNA decay, cell malfunction, and pathogenesis [4, 6].
Increasingly, abnormalities in tRNA expression, modification, and charging are linked to cancer [5, 7], cardiovascular [8, 9], neurodegenerative [10], and viral infectious [11] diseases.
Despite their vital importance, profiling tRNA expression, modification, and charging had been technically difficult in the past due to the highly stable tRNA fold structure and heavy tRNA modifications that hinder traditional small RNA sequencing library construction, resulting in poor cDNA yields, biased fragment coverage, and low quantitative accuracy. Arraystar tRNA Charging Seq is the most advanced technology to overcome all the difficulties to simultaneously profile tRNA expression, tRNA modifications, and tRNA charging.
From total RNA to tRNA expression, modification, and charging profiles
RNA quality and quantity assessment before the project proceeds, with small-RNA-retaining purification.
A thermostable reverse transcriptase synthesizes full-length cDNA for tRNAs at high efficiency, overcoming reverse transcription blocks by tRNA modifications.
Construction and high-throughput sequencing of the tRNA cDNA library.
Base misincorporations induced by modifications pinpoint modified positions; 3′-CCA end oxidation/β-elimination analysis measures amino acid charged or uncharged status.
Differential analyses with detailed annotations for tRNA expression, modification, and charging, with publication-ready graphics and visualization.
A wealth of tRNA multi-omics analyses included in every project
tRNA Charging Seq provides a wealth of tRNA multi-omics data with common analyses (e.g. differential analyses) and detailed annotations, for comprehensive insights into the tRNAs. Integrative analyses are also available with other Arraystar services: mRNA-seq, tRF&tiRNA-Seq, Arraystar Small RNA Microarray, and Arraystar Small RNA Modification Microarray.
tRNA multi-omics applications across biology and disease
Valine aminoacyl-tRNA synthetase promotes therapy resistance in melanoma (Nat Cell Biol, 2024); tRNA profiles inform drug resistance and tumor progression studies.
Glutamyl-prolyl-tRNA synthetase regulates proline-rich pro-fibrotic protein synthesis during cardiac fibrosis (Circ Res, 2020).
AIMP3 maintains cardiac homeostasis by regulating the editing activity of methionyl-tRNA synthetase (Nat Cardiovasc Res, 2025).
High-fidelity and differential nonsense suppression in live cells and a frontotemporal dementia allele with human transfer RNAs (Nucleic Acids Res, 2025).
Anticodon engineered transfer RNA inhibits hepatitis B virus replication by promoting degradation of core protein (Adv Sci, 2025).
tRNA expression, modification, and charging profiles directly regulate translation efficiency and accuracy by codon preferences, linking to translatomics studies.
Official Arraystar sample submission requirements for tRNA Charging Seq
Common questions about tRNA Charging Seq
Key References for tRNA Charging Seq
Arraystar tRNA Charging Seq delivers tRNA expression, modification, and charging profiles in a single assay — get a quote and a project timeline tailored to your study.