Barcoding intracellular reverse transcription enables high-throughput phenotype-coupled T cell receptor analyses.

ABSTRACT: Assays linking cellular phenotypes with T cell or B cell antigen receptor sequences are crucial for characterizing adaptive immune responses. Existing methodologies are limited by low sample throughput and high cost. Here, we present INtraCEllular Reverse Transcription with Sorting and sequencing (INCERTS), an approach that combines molecular indexing of receptor repertoires within intact cells and fluorescence-activated cell sorting (FACS). We demonstrate that INCERTS enables efficient processing of millions of cells from pooled human peripheral blood mononuclear cell (PBMC) samples while retaining robust association between T cell receptor (TCR) sequences and cellular phenotypes. We used INCERTS to discover antigen-specific TCRs from patients with cancer immunized with a novel mutant KRA

SUBMITTER: Jayaraman S 

PROVIDER: S-EPMC10626196 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature