Supplementary Figure 9: Transcriptional effects of CRISPR CREB-Zfp189 interactions in PFC.
From: Stress resilience is promoted by a Zfp189-driven transcriptional network in prefrontal cortex
a-b) GOBP pathways affected by dCas9-CREBS133D combined with Zfp189-sgRNA in PFC in (a) mice exposed to social defeat and (b) unstressed controls. (DEG threshold = p < 0.05, FC > log2|0.2|, Asterisk = FDR q < 0.05) c) Overlap for PFC DEGs (p < 0.05, log2FC > |0.2|) resulting from Zfp189-targeted dCas9-CREBS133D compared to NT-dCas9-CREBS133D in unstressed controls. d) Pink module genes differentially expressed in unstressed controls. e) On and off-target CRISPR gene regulation in RNA-seq datasets. Expression of the targeted Zfp189 gene is significantly up-regulated (p < 0.05, FC > log2|0.2|) in both defeated and un-stressed mice. The other 49 genes analyzed were those in closest proximity to other genomic regions with closest homology to the Zfp189-sgRNA used. No genomic region contains fewer than 3 mismatches with this sgRNA. Only one of these predicted off-target sites is affected by dCas9-CREBS133D plus Zfp189-sgRNA, but this regulation was only seen in defeated mice, suggesting that this is not an off-target effect of the CRISPR tools, but rather an effect of the defeat per se.
