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Exosomal tRFs & Liquid Biopsy Solution — From Biofluid tRF/tiRNA Discovery to Non-Invasive Biomarkers

tRNA and tRF&tiRNA populations are highly enriched in biofluids, much more so than microRNAs, making them excellent biomolecules for biomarkers. Arraystar's solution profiles tRFs and tiRNAs in serum, plasma, and exosomes by tRF&tiRNA-seq, then validates candidate biomarkers with the nrStar™ tRF&tiRNA PCR Array.

Biofluid-compatible · exosome workflows · discovery to validation

Overview

Non-invasive biomarker discovery from biofluid RNA

The composition and abundance of tRFs&tiRNAs are highly dependent on the cell type and disease condition, making them excellent biomolecules for biomarkers [1]. Also, tRNA and tRF&tiRNA populations are highly enriched in biofluids, much more so than microRNAs [2-3]. Exosome-derived tRFs have been reported in extracellular vesicles from plasma, serum, and other biofluids [4].

Arraystar's exosomal tRF & liquid biopsy solution starts with tRF&tiRNA-seq on serum, plasma, or exosome-derived RNA to discover differentially expressed tRFs and tiRNAs. Candidate biomarkers are then validated across cohorts using the nrStar™ tRF&tiRNA PCR Array or pre-designed tRF/tiRNA primers, with the rtStar™ tRF&tiRNA Pretreatment & First-Strand cDNA Synthesis Kit ensuring accurate detection.

What is the exosomal tRFs & liquid biopsy solution?

This solution profiles tRNA-derived fragments (tRFs) and tRNA halves (tiRNAs) in biofluids such as serum, plasma, and exosomes. It combines tRF&tiRNA-seq for discovery with tRF&tiRNA PCR arrays and pre-designed primers for validation, supporting non-invasive biomarker research.

Service at a Glance

Liquid biopsy services — from biofluid RNA to validated tRF/tiRNA biomarkers

Service NameCoveragePrice
tRF&tiRNA SequencingGenome-wide tRF/tiRNA discovery from biofluid RNA
nrStar™ tRF&tiRNA PCR Array (H/M)185 human / 88 mouse prevalent tRFs & tiRNAs
Pre-designed tiRNA&tRF Primer SetIndividual tRF/tiRNA quantification

Benefits

Why choose Arraystar for liquid biopsy research

🩸

Biofluid-Ready

Workflows for serum, plasma, and exosome-derived RNA, where tRFs/tiRNAs are highly enriched.

🔬

Genome-Wide Discovery

tRF&tiRNA-seq profiles the full tRF/tiRNA landscape in biofluid samples.

✅

High-Throughput Validation

PCR arrays and pre-designed primers validate candidates across cohorts in hours.

🎯

Precursor Discrimination

Adaptor-based assays distinguish tRFs/tiRNAs from tRNA and pre-tRNA precursors.

🧬

Accurate Detection

rtStar™ pretreatment removes internal and terminal modifications for accurate qPCR.

📊

Focused Bioinformatics

Differential expression, subtype analysis, and publication-quality graphics.

Background

tRFs and tiRNAs in biofluids

tRFs and tiRNAs, generated through precise biogenesis processes from tRNA, perform many biological functions as small noncoding RNAs and are associated with many diseases and conditions (Fig. 1). The composition and abundance of tRFs&tiRNAs are highly dependent on the cell type and disease condition, making them excellent biomolecules for biomarkers [1].

Also, tRNA and tRF&tiRNA populations are highly enriched in biofluids, much more so than microRNAs [2-3]. Extracellular vesicles carry biologically active tRFs and tiRNAs: maternal circulating syncytiotrophoblast-derived extracellular vesicles contain biologically active 5′ tRNA halves [4], and tRNA-derived fragments secreted by extracellular vesicles have diagnostic value in hypopharyngeal carcinoma [5].

Figure 1. tRF&tiRNA functions and association with diseases.
Figure 1. tRF&tiRNA functions and association with diseases.

Background References

  1. Telonis AG, et al. Dissecting tRNA-derived fragment complexities using personalized transcriptomes reveals novel fragment classes and unexpected dependencies. Oncotarget, 2015. PMID: 26325506
  2. Schageman J, et al. The complete exosome workflow solution: from isolation to characterization of RNA cargo. BioMed Research International, 2013. PMID: 24205503
  3. Dhahbi JM, et al. 5′ tRNA halves are present as abundant complexes in serum, concentrated in blood cells, and modulated by aging and calorie restriction. BMC Genomics, 2013. PMID: 23638709
  4. Cooke WR, et al. Maternal circulating syncytiotrophoblast-derived extracellular vesicles contain biologically active 5′ tRNA halves. Biochemical and Biophysical Research Communications, 2019. PMID: 31405560

Solution Workflow — Building Your Evidence Chain

From biofluid samples to validated liquid biopsy biomarkers

1

Biofluid Collection & RNA QC

Serum, plasma, or exosome samples are collected per official guidance; RNA is purified with small-RNA retention and quality checked.

2

Genome-Wide Discovery

tRF&tiRNA-seq profiles differentially expressed tRFs and tiRNAs from biofluid RNA with precise annotation.

3

Bioinformatics Prioritization

Differential expression, subtype analysis, and statistics prioritize candidate biomarkers.

4

Cohort Validation

The nrStar™ tRF&tiRNA PCR Array or pre-designed primers validate candidates across many samples.

5

Integrative Interpretation

Results are interpreted with clinical variables and, where relevant, tRNA modification data.

Bioinformatics & Deliverables

Deliverables for liquid biopsy projects

Sequencing projects deliver a wealth of tRF&tiRNA-focused bioinformatics; array projects deliver ready-to-analyze qPCR data with analysis tools. As a bonus, microRNA expression analysis is included in tRF&tiRNA-seq projects.

Standard Deliverables

Research Applications

Liquid biopsy research applications

Cancer Diagnostics

tRNA-derived fragments secreted by extracellular vesicles have diagnostic value in hypopharyngeal carcinoma (OncoTargets Ther, 2021).

Cardiovascular Disease

Differentially expressed tRNA-derived fragments in primary cardiomyocytes stimulated by high glucose (Front Endocrinol, 2023).

Ocular Complications

tRF-1020 ameliorates diabetes-induced retinal microvascular complications (J Cell Mol Med, 2022).

Placental Biology

Maternal circulating syncytiotrophoblast-derived extracellular vesicles contain biologically active 5′ tRNA halves (BBRC, 2019).

Prostate Cancer

A comprehensive repertoire of tRNA-derived fragments in prostate cancer supports biomarker discovery (Oncotarget, 2016).

Choosing the Right Strategy for Your Study

Method selection depends on your discovery stage and sample type

DimensiontRF&tiRNA SequencingnrStar™ tRF&tiRNA PCR ArrayPre-designed Primer Sets
Profiling scopeGenome-wide tRFs & tiRNAs with precise annotation185 human / 88 mouse prevalent tRFs & tiRNAsIndividual tRFs/tiRNAs of interest
Best forDiscovery in biofluid samplesHigh-throughput cohort validationTargeted quantification
Sample throughputLow to moderateHigh (384-well plates, hours)Flexible
RNA input> 1 µg> 5 µg> 5 µg

Sample Requirements

Official Arraystar sample submission requirements for liquid biopsy projects

RNA Amount & Quality

  • Biofluid input: serum or plasma 1-2 mL (0.3 mL/aliquot) without IP/pretreatment, 2-5 mL with IP/pretreatment; exosome isolation 2 mL. Whole blood 1-2 mL (0.3 mL/aliquot).
  • Total RNA input: > 1 µg for tRF&tiRNA-seq; > 5 µg for PCR array projects (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 tRF&tiRNA are < 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: RNAs well known to be degraded or fragmented (e.g. serum/plasma/exosomal RNA) are not checked for RNA integrity.

Shipping Instructions

  • Ship RNA in nuclease-free water (> 20 ng/µL), freeze-dried, or in ethanol; store at −80 °C or in liquid nitrogen.
  • Follow the official serum/plasma collection guidance (e.g. 0.3 mL aliquots) to preserve small RNA content.
  • 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.

FAQ

Common questions about exosomal tRFs & liquid biopsy

Why are tRFs/tiRNAs attractive for liquid biopsy?
tRNA and tRF&tiRNA populations are highly enriched in biofluids, much more so than microRNAs, and their composition and abundance depend on the cell type and disease condition. This makes them excellent biomolecules for biomarkers, including in serum, plasma, and exosome-derived RNA.
Which sample types are supported?
The solution supports serum, plasma, whole blood, and exosome-derived RNA, following the official biofluid collection guidance (e.g. serum/plasma 1-2 mL without pretreatment, 2-5 mL with pretreatment, 2 mL for exosome isolation). RNAs known to be degraded or fragmented are not checked for integrity.
What is the minimum amount of RNA required?
Recommended minimums are more than 1 µg of total RNA for tRF&tiRNA-seq and more than 5 µg for PCR array projects — official Arraystar recommended minimums for the entire experiment in a single attempt, including sample QC. Supplying twice the recommended minimum helps avoid project delays.
How are tRFs/tiRNAs distinguished from tRNA precursors?
The nrStar™ tRF&tiRNA PCR Array system introduces 3′ and 5′ adaptors to tRF/tiRNA fragments; forward and reverse primers targeting the adaptor junctions effectively discriminate tRFs/tiRNAs from tRNA and pre-tRNA precursors, giving reliable and accurate detection of tRF/tiRNA expression in biofluid samples.
What bioinformatics analyses are included?
tRF&tiRNA-seq projects include differential expression profiling with detailed annotation, subtype pie charts, and focused tRF/tiRNA statistics, plus a bonus microRNA expression analysis. Array projects include ready-to-analyze qPCR data with analysis tools and quality controls for validation studies across cohorts and biofluids.
Can the solution be combined with other Arraystar services?
Yes. The rtStar™ tRF&tiRNA Pretreatment & First-Strand cDNA Synthesis Kit ensures accurate qPCR detection, and results can be integrated with tRNA sequencing, tRNA modification analysis, and other services in Arraystar's tRNA research portfolio for comprehensive liquid biopsy studies across disease areas.

Selected Publications

Featured Client Publications in Exosomal tRF & Liquid Biopsy Research

  1. Xi J, et al. Expression and Diagnostic Value of tRNA-Derived Fragments Secreted by Extracellular Vesicles in Hypopharyngeal Carcinoma. OncoTargets and Therapy, 2021. PMID: 34285510
  2. Cooke WR, et al. Maternal circulating syncytiotrophoblast-derived extracellular vesicles contain biologically active 5′ tRNA halves. Biochemical and Biophysical Research Communications, 2019. PMID: 31405560
  3. Ma C, et al. tRNA-derived fragment tRF-1020 ameliorates diabetes-induced retinal microvascular complications. Journal of Cellular and Molecular Medicine, 2022. PMID: 36128646
  4. Zhao Y, et al. Differentially expressed tRNA-derived fragments and their roles in primary cardiomyocytes stimulated by high glucose. Frontiers in Endocrinology, 2023. PMID: 36714586
  5. Olvedy M, et al. A comprehensive repertoire of tRNA-derived fragments in prostate cancer. Oncotarget, 2016. PMID: 27015120
  6. Dhahbi JM, et al. 5′ tRNA halves are present as abundant complexes in serum, concentrated in blood cells, and modulated by aging and calorie restriction. BMC Genomics, 2013. PMID: 23638709

Ready to Profile tRFs in Biofluids and Exosomes?

Arraystar's exosomal tRFs & liquid biopsy solution combines tRF&tiRNA-seq discovery with PCR array validation — get a quote and a project timeline tailored to your study.