Chunk #9 — Results — The Efficacy of dCas9-Mediated Transcript Activation Varies Extensively between Genes and Is Not Necessarily Consistent between Unique Donors
First, transcriptional modulation of KCTD13 and TOAK2 was evaluated in NPCs. Across hiPSC-NPC lines from three unique donors, following the transduction of different lentiviruses encoding three distinct gRNAs, two KCTD13 gRNAs (2 and 3) significantly increased KCTD13 in dCas9−VP64 NPCs from two of the three individuals tested (gRNA 2: C1, 2.22-fold, p ≤ 0.01; C2, 1.87-fold, p ≤ 0.01; n = 3 each; gRNA 3: C1, 1.85-fold, p ≤ 0.05; C2, 1.48-fold, p ≤ 0.05; n = 3 each; antibiotic selection for dCas9−VP64) (Figure 2A). Activation of TOAK2 by gRNAs targeted to three different locations upstream of the TSS failed to increase expression in dCas9−VP64 NPCs across both individuals tested (antibiotic selection for dCas9−VP64) (Figure 2B).
