Achieve functional knockout in human primary T-cells

T cell receptor knockout using TrueGuide Synthetic gRNAs. Human T cells were isolated and activated using Dynabeads magnetic beads (Cat. Nos. 11344D and 11141D, respectively) and then transfected with TrueCut Cas9 Protein v2 and TrueGuide Synthetic gRNA for T cell receptor alpha (TRAC) or beta (TRBC) regions using the Neon Transfection System. Following transfection, editing efficiency was measured by the GeneArt Cleavage Detection assay (a) or by measuring % T cell receptor negative (TCR–) cells using the Attune NxT Flow Cytometer (b). Cells analyzed by flow cytometry were stained with a T cell receptor–specific antibody conjugated to PE (c).