Feb 11, 2020

Very rapid cloning, expression and identifying specificity of T-cell receptors for T-cell engineering

PloS One
Shan ZongHiroki Torikai

Abstract

Neoantigens can be predicted and in some cases identified using the data obtained from the whole exome sequencing and transcriptome sequencing of tumor cells. These sequencing data can be coupled with single-cell RNA sequencing for the direct interrogation of the transcriptome, surfaceome, and pairing of αβ T-cell receptors (TCRαβ) from hundreds of single T cells. Using these 2 large datasets, we established a platform for identifying antigens recognized by TCRαβs obtained from single T cells. Our approach is based on the rapid expression of cloned TCRαβ genes as Sleeping Beauty transposons and the determination of the introduced TCRαβs' antigen specificity and avidity using a reporter cell line. The platform enables the very rapid identification of tumor-reactive TCRs for the bioengineering of T cells with redirected specificity.

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Mentioned in this Paper

Gene Cloning
Biomedical Engineering Field
T-Cell Receptor
Neoplastic Cell
C-reactive protein
DNA Transposable Elements
Transcriptome
Antigens
Determination Aspects
Whole Exome Sequencing

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