A method for integrating cellular phenotype with T cell receptor analysis
JHU Ref #: C17820
Value Proposition:
· Paired TCR sequencing and phenotypic analysis
· High throughput and resolution
· Enables pooling of samples
Unmet Need
Characterization of the adaptive immune response has been advanced by single cell RNA-sequencing (scRNA-seq) technology. Profiling immune cells and their respective receptors better informs target antigens and the function of the cell to develop precise immunotherapies. Cellular phenotype can be determined by flow cytometric sorting of cells for bulk repertoire sequencing. However, bulk-seq is limited in phenotypic information. On the other hand, scRNA-seq provides paired alpha and beta TCR sequences for identifying antigen specificity. scRNA-seq is limited by throughout and the cost to scale is prohibitive. Therefore, there is a strong need for the integration of the two methods for high resolution phenotypic and paired TCR sequences information to identify key features of the adaptive immune response.
Technology Description
Researchers at Johns Hopkins have developed a method for pairing TCR sequencing to phenotypic analysis. The technology combines indexing of receptor repertoires within cells and fluorescence-activated cell sorting (FACS) by using sample specific barcodes. The method, called INtraceCEllular Reverse Transcription with Sorting and sequencing (INCERTS), enables pooling of samples for increased feasibility and throughput.
Stage of Development
INCERTS methodology validated with human PBMC samples.
Data Availability
· Data available upon request.
Publication
Barcoding intracellular reverse transcription enables high-throughput phenotype-coupled T cell receptor analyses
Jayaraman, Sahana et al. Cell Reports Methods, Volume 3, Issue 10, 100600
PCT WO2025/072917 BARCODING INTRACELLULAR REVERSE TRANSCRIPTION ENABLES HIGH-THROUGHPUT ANALYSIS OF RNA SEQUENCES
