Fig. 5.

Editing the γ-globin transgenes eliminates bone marrow engraftment of Berkeley mouse HSCs. (A) Experimental strategy. Lin⁻ HSPCs from Berkeley mice (CD45.2) were transfected with RNP targeting HBG1/HBG2 or Rosa26, transplanted into irradiated C57Bl/6 (CD45.1) hosts and analyzed serially. (B) Kaplan–Meier curves showing survival of irradiated recipients after transplantation of edited HSPCs. (C) Engraftment of HBG1/HBG2-edited or Rosa26-edited Berkeley mouse donor HSPCs over time, as determined by the fraction of circulating CD45.2⁺ mononuclear cells in recipient mice. (D) Frequency of on-target indels at the HBG or Rosa26 loci in blood mononuclear cells at the indicated times following transplantation. Graphs show mean±s.e.m. of three biological replicate experiments with n=10 HBG-edited, n=6 Rosa26-edited and n=10 control (Cas9 only-transfected) mice. ns, not significant; **P<0.01, ***P<0.001, ****P<0.0001, by unpaired, two-tailed Student's t-test, survival calculations Log-rank (Mantel–Cox) test.