Fig. 3
From: Somatic genome editing with the RCAS-TVA-CRISPR-Cas9 system for precision tumor modeling
Ubiquitous Cas9 expression in TVA-Cas9 adult mice does not induce a robust immune response. a Left panels: Flow cytometry analysis for the specified markers in blood and spleen of Ntv-a; LSL-Cas9; hUBC-CreERT2 mice of the indicated genotype. Four-week-old mice were treated with tamoxifen in the food for 5 consecutive weeks. Right panels: representative flow cytometry plots, with the gating strategy used for the analysis. Student’s t test: ***P < 0.001, **P < 0.005, *P < 0.05. The upper and lower hinges correspond to the first and third quartiles (the 25th and 75th percentiles). The upper whisker extends from the hinge to the highest value that is within 1.5 × IQR of the hinge, where IQR is the inter-quartile range. The lower whisker extends from the hinge to the lowest value within 1.5 × IQR of the hinge. b Left panels: Flow cytometry analysis for lymphocytes, macrophages and microglia of the brain of the mice in a. Right panels: representative flow cytometry plots, with the gating strategy used for the analysis. c Table summarizing the injections performed in the Ntv-a; LSL-Cas9; hUBC-CreERT2 adult mice. Mice injected with RCAS-PDGFB + RCAS-Trp53-gRNA and treated with tamoxifen to induce Cas9 expression, develop high-grade gliomas. d H&E and Cas9 IHCs of RCAS-PDGFB + RCAS-Trp53-gRNA tumors for the indicated treatment. High-grade glioma features and Cas9 expression are present only in the tamoxifen-treated mice. Scale bars: H&E 100 μm; IHCs 50 μm
