HTGTS-TCR-Seq for Profiling of Mouse and Human T-Cell Receptor α and β Gene Rearrangements and Diversity.
Level 5 - mechanism / opinion, no new human data
Preclinical laboratory method development and sequencing validation in mouse models and human cellular samples
PubMed 41144713 · doi:10.1002/advs.202509497
What was done
Researchers developed HTGTS-TCR-seq (high-throughput genome-wide translocation sequencing-based TCR sequencing), a method using a set of 3–5 J or V region primers to enrich for V(D)J recombination products and profile Tcra and Tcrb rearrangements. The technique was tested on wild-type mouse thymocytes across developmental stages, young and aged mouse T cells, Wapl-knockout preselection double-positive thymocytes, and human peripheral T cells.
What was found
The abstract reports no numerical data, error rates, or statistical values. It reports that the assay detected both productive and nonproductive rearrangements, stage-specific V and J usage, and age-associated repertoire alterations in mice. In Wapl-knockout thymocytes, it identified a cell division-independent role for WAPL in Tcra rearrangement. In human peripheral T cells, it observed conserved CDR3 characteristics and subset-specific Vbeta usage.
Why it matters
The method offers a cost-effective, low-primer alternative to multiplex PCR and 5'RACE for capturing comprehensive TCR alpha and beta repertoires, including nonproductive alleles useful for developmental tracking.
Limits
The abstract reports no quantitative benchmarking metrics (such as capture efficiency, sequencing error rates, or sensitivity thresholds) directly comparing the method against existing 5'RACE or multiplex platforms. Sample sizes for mouse cohorts and human donors are not reported.