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. 2023 May 22;10(22):2301802. doi: 10.1002/advs.202301802

Figure 4.

Figure 4

CRISPR‐dCas9‐KRAB induces sustained silencing of ZEB1 in vivo. Representative images of the resected dCas9‐KRAB‐edited tumors versus controls (Figure 3) by immunofluorescence for the detection of (A) dCas9 (green) and (B) ZEB1 (red). MDA‐MB‐231‐luc tumors either untransduced (Wild type) or transduced with dCas9‐KRAB in absence of gRNA (No gRNA), or expressing All gRNA were analyzed at day 32 post‐implantation of the cells. Hematoxylin & Eosin (H&E) staining of serial sections is indicated to illustrate cellularity. (C) Box and whisker plot highlighting the percent staining intensity of dCas9 and ZEB1 in tumors extract at day 32 relative to untransduced (Wild type). (D) Gene expression analyses by qRT‐PCR to assess the regulation of the pro‐epithelial miR‐200 family members in vivo, with tumors resected at day 32 post‐implantation. ns = non‐significant, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and ****p ≤ 0.0001; miR: microRNA.