Fig. 2
From: Multifunctional CRISPR-Cas9 with engineered immunosilenced human T cell epitopes
SpCas9 immunodominant epitope-specific CD8+T cell recognition is abolished after anchor residue mutation. a Epitope β-specific CD8+T cell response detected using β-specific pentamer in PBMCs stimulated with peptide β-pulsed antigen presenting cells. b The percentage of CD8+ pentamerβ+T cells was reduced to 0.3% when healthy donor B cell APCs were pulsed with the mutated peptide β2. c Positions, sequences, and IEDB HLA binding percentile rank of epitopes α and β before and after mutation of the anchor (2nd and/or 9th) residues. Sb, normalized binding score; Si, normalized immunogenicity score. d Representative IFN-γ ELISpot assay in triplicate wells comparing T cell reactivity to wild type or mutated epitopes α and β. These results are representative of 12 donors and two independent replicates (data from all 12 donors are shown in Supplementary Fig. 1). e, f IFN-γ ELISpot comparing T cell reactivity to APCs expressing WT or modified Cas9 proteins. APCs expressing FluM1 were used as a positive control. APCs expressing GAPDH or spiked with peptide α2 were used as negative controls. Data represent mean ± SEM of 5 replicates (right). Statistical analysis was performed post hoc and results are exploratory. Source data are available in the Source Data file
