An HD-Ad5/35⁺⁺ vector for multiplex mutagenesis in human erythroid cells. (A) Design of the 3 adenoviral vectors used in downstream experiments. (B) HbF expression in bulk-transduced HUDEP-2 cells after transduction with the different Ad-CRISPRs. All samples were transduced with the same MOI. (C) Representative HbF FACS dot plots. (D) γ-globin expression as evaluated by HPLC. (E) HbF expression presented as mean fluorescent intensity (MFI) in single-cell clones derived from Ad-BCL11A–, Ad-HBG-115–, or Ad-BCL11A + HBG-115–transduced cells. Values are represented as means ± SEM. ***P ≤ .0001, **P ≤ .001, *P ≤ .05 (unpaired Student t test).