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Eur J Immunol. 2019 Apr 24. doi: 10.1002/eji.201848030. [Epub ahead of print]

High-throughput single-cell sequencing of paired TCRα and TCRβ genes for the direct expression-cloning and functional analysis of murine T-cell receptors.

Author information

1
Department of B Cell Immunology, German Cancer Research Center, Heidelberg, Germany.

Abstract

Precise clonal and functional assessments of the T cell receptor (TCR) repertoire diversity require paired TCRα and TCRβ gene sequence information at monoclonal level. However, available single-cell strategies are typically limited in throughput and often do not provide full-length DNA templates for direct gene cloning. Here, we describe a high-throughput strategy for the unbiased amplification and automated sequence analysis of paired TCRα and TCRβ genes from primary mouse T cells. The platform links cell phenotype and TCR gene sequence information at single-cell level. Furthermore, it enables direct functional analyses through the efficient cloning of both genes and the generation of stable TCR expressing cell lines. This highly efficient workflow is a powerful tool to determine the diversity and quality of the murine T-cell repertoire in various settings, for example in vaccine development, infectious diseases, autoimmunity, or cancer.

KEYWORDS:

Expression cloning; Functional TCR analysis; Single cell; T cell receptor

PMID:
31017295
DOI:
10.1002/eji.201848030

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