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RNA Modifying Enzyme Target & Mechanism Solution — From Enzyme Expression to Modification Site and Function

tRNA modifications are dynamically regulated and catalyzed by tRNA modification enzymes; mutations or dysregulation of these modifiers are linked to diseases such as microcephaly and type 2 diabetes. Arraystar's solution profiles enzyme expression with the NuRNA™ tRNA Modification Enzymes PCR Array, quantifies global modification levels by LC-MS, and maps modification sites at base resolution.

Enzyme expression · global modification quantification · single-base site mapping

Overview

Three complementary layers of evidence for enzyme function

Chemical modifications are dynamically regulated and catalyzed by tRNA modification enzymes [1]. Mutations or dysregulation of tRNA modifiers have been associated with diseases, for example, mutations in cytosine-5 RNA methyltransferase (NSUN2) with microcephaly [2] and t6A methylthiolation enzymes with type 2 diabetes [3]. How tRNA modifications are regulated by tRNA modification enzymes is in need of study.

Arraystar's RNA modifying enzyme solution combines three layers of evidence: the NuRNA™ Human tRNA Modification Enzymes PCR Array profiles the expression of 85 enzymes and protein factors; LC-MS based tRNA modification analysis quantifies 55 nucleoside modifications globally; and tRNA Modification Seq, m7G TRAC-Seq, and m3C HAC-Seq map specific modifications at single-base resolution to confirm enzyme targets.

What is the RNA modifying enzyme target & mechanism solution?

This solution links tRNA modification enzymes to their targets and mechanisms by profiling enzyme expression (NuRNA™ PCR Array), quantifying global modification levels (LC-MS), and mapping modification sites at base resolution (Modification Seq, TRAC-Seq, HAC-Seq), supporting disease mechanism studies.

Service at a Glance

Enzyme mechanism services — combine expression profiling, global quantification, and site mapping

Service NameProfiling LayerPrice
NuRNA™ tRNA Modification Enzymes PCR ArrayExpression of 85 tRNA modification enzymes
LC-MS Based tRNA Modification Analysis55 nucleoside modifications (global profile)
tRNA Modification Seq – m1A, m3C, m1G, m2,2GSingle-base resolution for 4 methylation types
m7G TRAC-Seqm7G sites at single-nucleotide resolution
m3C HAC-Seqm3C sites at single-nucleotide resolution

Benefits

Why choose Arraystar for enzyme mechanism studies

📚

85 Enzyme Panel

The first commercial PCR panel profiling 85 validated or predicted tRNA modification enzymes and protein factors.

📊

Global Modification Profile

LC-MS quantifies 55 nucleoside modifications to capture the overall modification landscape.

🎯

Single-Base Site Mapping

Confirm enzyme targets at nucleotide-level accuracy with chemical-specific methods.

Rigorous Validation

Array assays validated across many tissues and cell lines with full quality controls.

🧬

Disease Relevance

Enzyme–disease links including NSUN2–microcephaly and CDKAL1–type 2 diabetes.

🔗

Integrated Evidence Chain

Expression, quantification, and site mapping combined in one research program.

Background

tRNA modification enzymes and disease

tRNAs are heavily decorated posttranscriptionally with numerous chemical modifications. The modifications are essential for shaping up, fine tuning, and regulating all aspects of tRNA functioning, such as folding, stability, and decoding. These chemical modifications are dynamically regulated and catalyzed by tRNA modification enzymes [1].

Mutations or dysregulation of tRNA modifiers have been associated with diseases, for example, mutations in cytosine-5 RNA methyltransferase (NSUN2) with microcephaly [2] and t6A methylthiolation enzymes with type 2 diabetes [3]. The emerging importance of tRNA modifications in diseases calls for additional work [4, 5].

Figure 1. Human tRNA modifications and tRNA modification enzymes (in parentheses).
Figure 1. Human tRNA modifications and tRNA modification enzymes (in parentheses).

Background References

  1. El Yacoubi B, et al. Biosynthesis and function of posttranscriptional modifications of transfer RNAs. Annual Review of Genetics, 2012. PMID: 22905870
  2. Blanco S, et al. Aberrant methylation of tRNAs links cellular stress to neuro-developmental disorders. The EMBO Journal, 2014. PMID: 25063673
  3. Zhou B, et al. Identification of a splicing variant that regulates type 2 diabetes risk factor CDKAL1 level by a coding-independent mechanism in human. Human Molecular Genetics, 2014. PMID: 24760768
  4. Kirchner S, Ignatova Z. Emerging roles of tRNA in adaptive translation, signalling dynamics and disease. Nature Reviews Genetics, 2015. PMID: 25534324
  5. Zhang X, et al. Small RNA Modifications: Integral to Function and Disease. Trends in Molecular Medicine, 2016. PMID: 27840066

Solution Workflow — Building Your Evidence Chain

From samples to enzyme–modification–disease links

1

Sample & RNA QC

RNA quality and quantity assessment before the project proceeds, with small-RNA-retaining purification.

2

Enzyme Expression Profiling

The NuRNA™ tRNA Modification Enzymes PCR Array profiles the expression of 85 enzymes and protein factors.

3

Global Modification Quantification

LC-MS quantifies 55 nucleoside modifications to capture the global modification profile of the samples.

4

Base-Resolution Site Mapping

Select the sequencing method by target: Modification Seq (m1A, m3C, m1G, m2,2G), TRAC-Seq (m7G), or HAC-Seq (m3C).

5

Integrative Interpretation

Correlate enzyme expression changes with modification levels and sites to identify targets and mechanisms.

Bioinformatics & Deliverables

Deliverables for enzyme mechanism projects

Each service includes detailed bioinformatics. Array projects deliver expression data with quality controls and analysis tools; LC-MS projects deliver raw and normalized peak data with chromatograms; sequencing projects deliver modification sites, levels, and motif analyses.

Standard Deliverables

Research Applications

Enzyme target and mechanism research across disease

Neurodevelopmental Disorders

Mutations in cytosine-5 RNA methyltransferase (NSUN2) are linked with microcephaly; aberrant tRNA methylation links cellular stress to neuro-developmental disorders (EMBO J, 2014).

Type 2 Diabetes

t6A methylthiolation enzymes (e.g. CDKAL1) are associated with type 2 diabetes risk (Hum Mol Genet, 2014).

Cancer

METTL1-mediated m7G modification of Arg-TCT tRNA drives oncogenic transformation; METTL1 overexpression drives tumorigenesis in multiple malignancies.

Cardiovascular Disease

Defects in tRNA modifications and modification enzymes are linked with cardiac conditions and mitochondrial-linked disorders.

Translation Regulation

Understand how modification enzymes shape tRNA folding, stability, and decoding through their targets.

Choosing the Right Strategy for Your Study

Method selection depends on the evidence layer you need

DimensionNuRNA™ PCR ArrayLC-MS AnalysisModification Seq / TRAC / HAC
Evidence layerEnzyme & protein factor expressionGlobal nucleoside modification levelsModification sites at base resolution
Best forScreening which enzymes changeQuantifying overall modification changesConfirming specific enzyme targets and sites
OutputExpression of 85 enzymes55 nucleoside modifications quantifiedSite positions, levels, and motifs
Recommended RNA> 5 µg10*–15 µg> 5 µg

Sample Requirements

Official Arraystar sample submission requirements for enzyme mechanism projects

RNA Amount & Quality

  • Total RNA input: > 5 µg for the NuRNA™ PCR Array and modification sequencing; 10*–15 µg for LC-MS analysis (official recommended minimums, including sample QC). Supply twice the recommended minimum to avoid project delays.
  • Purification: TRIzol / RNA precipitation or an RNA isolation kit. Because tRNA is < 200 nt, use a kit specified to retain small RNAs (e.g. Qiagen miRNeasy).
  • Concentration: > 20 ng/µL by Nanodrop; OD260/280 ~2.0 (acceptable 1.7–2.1); OD260/230 > 1.8.
  • Integrity: sharp 18S/28S rRNA bands by gel, or RIN > 7.0 by Bioanalyzer (serum/plasma/exosome/FFPE RNA exempt).
  • DNase treatment: required for qPCR projects; optional for sequencing and LC-MS.

Shipping Instructions

  • Ship RNA in nuclease-free water (> 20 ng/µL), freeze-dried, or in ethanol; store at −80 °C or in liquid nitrogen.
  • Use nuclease-free certified, screw-cap 1.5 mL microtubes; seal caps with Parafilm; place tubes in a plastic bag.
  • Use 10 kg dry ice as refrigerant; include a signed Project Form and the sample list.
  • Ship to: Arraystar Inc., 9430 Key West Avenue #128, Rockville, MD 20850, USA. Contact us before shipping.

FAQ

Common questions about RNA modifying enzyme target & mechanism studies

Which enzymes are covered by the NuRNA™ PCR Array?
The NuRNA™ Human tRNA Modification Enzymes PCR Array profiles the expression of 85 critical enzymes and protein factors involved in tRNA modifications, including ADAT1/2/3, ALKBH1/8, CDKAL1, METTL1, METTL2A/2B, NSUN2/6, PUS1/3/10, TRMT1, TRMT6/61A, and many more, each annotated with its corresponding modification.
How do I confirm the target of a specific enzyme?
First profile enzyme expression with the NuRNA™ PCR Array, then quantify global modification changes by LC-MS. To confirm the exact target sites, use tRNA Modification Seq (m1A, m3C, m1G, m2,2G), m7G TRAC-Seq, or m3C HAC-Seq, which map modifications at single-base resolution.
Why combine LC-MS with base-resolution sequencing?
LC-MS simultaneously quantifies 55 nucleoside modifications and characterizes the global modification profile of tRNAs, while sequencing methods localize individual modifications on specific tRNAs at single-base resolution. Together they connect overall modification changes with precise enzyme target sites, giving a complete picture of enzyme function and mechanism.
What is the minimum amount of RNA required?
Recommended minimums are more than 5 µg of total RNA for the PCR array and modification sequencing, and 10–15 µg for LC-MS analysis — official Arraystar recommended minimums for the entire experiment in a single attempt, including sample QC. Supplying twice the recommended minimum helps avoid delays.
Which diseases are linked to tRNA modification enzymes?
Mutations or dysregulation of tRNA modifiers are associated with diseases including microcephaly (NSUN2 mutations), type 2 diabetes (t6A methylthiolation enzymes such as CDKAL1), cancers (METTL1-mediated m7G), neurological syndromes, cardiac conditions, and mitochondrial-linked disorders, highlighting the disease relevance of enzyme studies.
What bioinformatics analyses are included?
Array projects include expression data with normalization references and quality controls; LC-MS projects include raw and normalized peak data with chromatograms; sequencing projects include modification sites, methylation levels, volcano plots, IGV read alignments, and sequence motif analyses, all with publication-ready graphics and visualization.

Selected Publications

Key References for tRNA Modification Enzyme Research

  1. Blanco S, et al. Aberrant methylation of tRNAs links cellular stress to neuro-developmental disorders. The EMBO Journal, 2014. PMID: 25063673
  2. Zhou B, et al. Identification of a splicing variant that regulates type 2 diabetes risk factor CDKAL1 level by a coding-independent mechanism in human. Human Molecular Genetics, 2014. PMID: 24760768
  3. El Yacoubi B, et al. Biosynthesis and function of posttranscriptional modifications of transfer RNAs. Annual Review of Genetics, 2012. PMID: 22905870
  4. Kirchner S, Ignatova Z. Emerging roles of tRNA in adaptive translation, signalling dynamics and disease. Nature Reviews Genetics, 2015. PMID: 25534324
  5. Zhang X, et al. Small RNA Modifications: Integral to Function and Disease. Trends in Molecular Medicine, 2016. PMID: 27840066
  6. Orellana EA, et al. METTL1-mediated m(7)G modification of Arg-TCT tRNA drives oncogenic transformation. Molecular Cell, 2021. PMID: 34352207

Ready to Link tRNA Modification Enzymes to Their Targets?

Arraystar combines enzyme expression profiling, global modification quantification, and single-base site mapping — get a quote and a project timeline tailored to your study.