< 0.0001/gRNA 3; 0.29-fold, p < 0.0001; C2, gRNA 2; 0.46-fold, p < 0.0001/gRNA 3; 0.28-fold, p < 0.0001; C3, gRNA 2; 0.33-fold, p < 0.0001/gRNA 3; 0.19-fold, p < 0.0001; n = 3 each; antibiotic selection for dCas9−KRAB) (Figure 3E). Efficacy of gRNA 2 in decreasing SNAP91 protein levels was confirmed by western blot (n = 3; antibiotic selection for dCas9−KRAB) (Figure S4B). Although gRNAs against CLCN3 were generally less efficacious, gRNA 3 produced significant effects (gRNA 3: C1, 0.85-fold, p < 0.01; C2, 0.74-fold, p < 0.01; C3, 0.82-fold, p < 0.01; n = 3 each; no antibiotic selection for dCas9−KRAB) in day 8 NGN2 neuron populations from all three NPC lines tested (Figure 3F).
