Supplementary Figure 2: AAV-delivered CRISPRa enhances the expression and presentation of targeted antigens.
From: Multiplexed activation of endogenous genes by CRISPRa elicits potent antitumor immunity
a, AAV-CRISPRa activation of multiple endogenous genes in a pooled manner. The transcript levels of Cd70, Cd80, Cd86 were quantified by qRT-PCR after transducing E0771-dCas9-VP64 cells with either AAV-Vector or AAV-sgRNA-miniLib (n = 3 cell infection replicates from one representative experiment). Two-sided unpaired t-test for sgRNAs vs. vector: Cd70, p < 0.0001; Cd80, p = 0.0005; Cd86, p < 0.0001. b, AAV-CRISPRa enhances the cell surface presentation of a model antigen in the form of peptide-MHC-I complexes. In cells transduced with lentivirus to express ovalbumin (Ova) under a PGK promoter, AAV-CRISPRa targeted to the PGK promoter augments the presentation of Ova-derived SIINFEKL-MHC-I complex on the cell surface. Quantitative analysis of geometric mean fluorescence intensity (MFI) of PE-SIINFEKL-H-2Kb on cells treated with AAV-Vector or AAV-sgRNAs. n = 6 cell infection replicates (SIINFEKL-H-2Kb staining in AAV-Vector), n = 6 (isotype in AAV-Vector), n = 6 (SIINFEKL-H-2Kb staining in AAV-sgRNAs), or n = 6 (isotype in AAV-sgRNAs) from four independent experiments. Two sided Mann-Whitney test: SIINFEKL-H-2Kb staining vs. isotype in AAV-Vector, p = 0.091; SIINFEKL-H-2Kb staining vs. isotype in AAV-sgRNAs, p = 0.0022; SIINFEKL-H-2Kb staining in AAV-sgRNAs vs. AAV-Vector, p = 0.0411. Error bars: All data in this figure are presented as mean ± s.e.m., with individual data points shown. Asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001.
