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

ABSTRACT: Precise clonal and functional assessments of the T cell receptor (TCR) repertoire diversity require paired TCR? and TCR? 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 TCR? and TCR? 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.

SUBMITTER: Ludwig J 

PROVIDER: S-EPMC6767390 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature