Figure 4
From: Hemophilia A ameliorated in mice by CRISPR-based in vivo genome editing of human Factor VIII
AAV dose-dependent expression of the targeted human BDD-F8 transgene ameliorated hemophilia A in mice. Adult F8KO or littermate WT control mice were injected without (Veh) or with the indicated total AAV doses of AAV8-SaCas9-sg1 and AAV8-BDD-F8 vectors, mixed at a 1:5 ratio. After 4 weeks, mouse plasma was collected and analyzed for FVIII protein level (A), FVIII activity (B) and activated partial thromboplastin time (aPTT, C). These parameters were assessed by FVIII ELISA, Chromogenix activity assay, and microplate aPTT assay, respectively, as described in Methods. (D) Tail vein bleeding time assays performed using filter paper, as described in Methods. Blood was collected into one spot of the filter paper every 4 min, with a total time of 28 min. Blood volume was calculated by a pre-defined standard curve of blood volume vs. spot area. Data are means ± SE. *P < 0.05, **P < 0.01 vs. F8KO mice injected with vehicle; #P > 0.3 vs. WT mice injected with vehicle; n = 3–5 mice per group. No coagulation (unlimited aPTT) is indicated by ∞.
