Sel scRNA-seq PT – Selective single cell transcriptomic analysis of rare cells for personalized therapy applications

We aim to develop a technology for selective single-cell transcriptomic (scRNA-seq) analysis of cells with a specific phenotype from a heterogeneous mixture. Cells will be encapsulated in picolitre droplets, with no more than one cell per droplet, together with single hydrogel beads carrying barcoded cDNA primers, with a unique barcode on each bead. Selective sequencing of only (rare) cells with the desired phenotype will be achieved by targeting single or multiple proteins expressed on the cell surface with DNA tagged antibodies which will trigger a molecular circuit called the DNA toolbox. In droplets containing cells with the desired phenotype, the molecular circuit will trigger selective release of barcoded cDNA primers from the hydrogel beads, ensuring that, after reverse transcription, barcoded cDNAs are only recovered from cells with the desired phenotype. Sequencing of the cDNAs will provide the transcriptomic landscape of only the target cells. The technology will be validated for scRNA-seq of infiltrating lymphocytes in solid tumor biopsies.