High-throughput single-cell sequencing of paired TCRα and TCRβ genes for the direct expression-cloning and functional analysis of murine T-cell receptors.
Expression cloning
Functional TCR analysis
Single cell
T cell receptor
Journal
European journal of immunology
ISSN: 1521-4141
Titre abrégé: Eur J Immunol
Pays: Germany
ID NLM: 1273201
Informations de publication
Date de publication:
08 2019
08 2019
Historique:
received:
20
12
2018
revised:
11
03
2019
accepted:
15
04
2019
pubmed:
25
4
2019
medline:
28
5
2020
entrez:
25
4
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31017295
doi: 10.1002/eji.201848030
pmc: PMC6767390
doi:
Substances chimiques
Receptors, Antigen, T-Cell, alpha-beta
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1269-1277Informations de copyright
© 2019 The Authors. European Journal of Immunology published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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