POKI-Seq in Primary Human T Cells. POKI-Seq in Primary Human T Cells

We adapted pooled non-viral targeted knock-ins to allow for simultaneous readouts of pooled knock-in barcode as well as RNA transcriptomes at the single cell level in primary human T cells. We used a library of 36 potentially therapeutic knock-in constructs each knocked into the TRAC locus along with a new TCR specificity to explore transcriptional changes in therapeutically relvant in vitro and in vivo environments. Overall design: Pooled CRISPR knock-in screen with single cell RNASeq readout (POKI-Seq) in primary human T cells. Dataset includes T cells from four donors, two donors each in two different experiemnts. The first experiment in vitro comparing TCR stimulation in the presense or absense of exogenous TGFB. The second experiment comparing in vivo TILs isolated from a solid tumor xenograft model against ex vivo expanded input cells.