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T cell receptor (TCR) technologies, including repertoire analyses and T cell engineering, are increasingly important in the clinical management of cellular immunity in cancer, transplantation, and other immune diseases. However, sensitive and reliable methods for repertoire analyses and TCR cloning are still lacking. Here, we report on SEQTR, a high-throughput approach to analyze human and mouse repertoires that is more sensitive, reproducible, and accurate as compared with commonly used assays, and thus more reliably captures the complexity of blood and tumor TCR repertoires. We also present a TCR cloning strategy to specifically amplify TCRs from T cell populations. Positioned downstream of single-cell or bulk TCR sequencing, it allows time- and cost-effective discovery, cloning, screening, and engineering of tumor-specific TCRs. Together, these methods will accelerate TCR repertoire analyses in discovery, translational, and clinical settings and permit fast TCR engineering for cellular therapies. © 2023 The Authors.

Citation

Raphael Genolet, Sara Bobisse, Johanna Chiffelle, Marion Arnaud, Rémy Petremand, Lise Queiroz, Alexandra Michel, Patrick Reichenbach, Julien Cesbron, Aymeric Auger, Petra Baumgaertner, Philippe Guillaume, Julien Schmidt, Melita Irving, Lana E Kandalaft, Daniel E Speiser, George Coukos, Alexandre Harari. TCR sequencing and cloning methods for repertoire analysis and isolation of tumor-reactive TCRs. Cell reports methods. 2023 Apr 24;3(4):100459

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PMID: 37159666

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