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Updated: Aug 29, 2025

Simultaneous Quantification of T-Cell Receptor Excision Circles TRECs and K-Deleting Recombination Excision Circles KRECs by Real-time PCR

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Tetramer-Associated T Cell Receptor Sequencing.

Michael Malone¹, Ke-Yue Ma², Shu-Qi Zhang³

¹Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA.

Methods in Molecular Biology (Clifton, N.J.)

|September 10, 2022

Summary

This summary is machine-generated.

Tetramer-associated TCR sequencing (TetTCR-Seq) links T cell receptor sequences to antigen specificity. This novel method enables rapid, cost-effective T cell repertoire analysis for therapeutic development.

Keywords:

DNA barcode In vitro transcription and translation TetTCR-Seq pMHC tetramer

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A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

Published on: March 22, 2012

Area of Science:

Immunology
Molecular Biology
Bioinformatics

Background:

Linking T cell receptor (TCR) sequences to their specific antigens is crucial for understanding T cell function and developing immunotherapies.
Current methods for associating TCR sequences with antigen specificity are often complex and labor-intensive.

Purpose of the Study:

To introduce and detail a novel method, tetramer-associated TCR sequencing (TetTCR-Seq), for efficiently linking TCR sequences with antigen specificity.
To provide a cost-effective and streamlined approach for high-throughput T cell repertoire analysis.

Main Methods:

TetTCR-Seq utilizes multiplexed DNA-barcoded peptide-MHC (pMHC) tetramers for simultaneous capture of antigen specificity and TCR sequences from single cells.
The method leverages in vitro transcription and translation (IVTT) to generate peptide libraries and DNA barcodes in parallel from a common template, reducing labor and cost.

Main Results:

TetTCR-Seq enables the simultaneous retrieval of antigen specificity and TCR sequences following single-cell sequencing.
The IVTT-based approach simplifies the generation of necessary reagents, contributing to reduced experimental costs and increased throughput.

Conclusions:

TetTCR-Seq is a powerful and innovative technology for surveying the T cell repertoire across numerous antigen specificities in a single experiment.
This method significantly advances the ability to perform large-scale T cell repertoire analysis, facilitating T cell biology research and the development of T cell-based therapeutics.