Extended Data Figure 5 |. Long-term multi-lineage reconstituting HSCs edited with 3xNLS-Cas9.

a-d, NBSGW mice were transplanted with 3xNLS-Cas9 RNP with MS-sgRNA-1617 edited healthy donor CD34⁺ HSPCs 2 h (day 0), 24 h (day 1) or 48 h (day 2) after electroporation. BM were collected 16 weeks after transplantation and analyzed by flow cytometry for human cell chimerism (a), multilineage reconstitution (b) or human erythroid cells (c) in BM, as well as indel frequencies determined by TIDE analysis (d). e-h, NBSGW mice were transplanted with 3xNLS-Cas9 RNP with MS-sgRNA-1617 edited healthy donor CD34⁺ HSPCs supplemented with 2%, 4% or 6% of glycerol for electroporation. BM were collected 16 weeks after transplantation and analyzed by flow cytometry for human cell chimerism (e), multilineage reconstitution (f) or human erythroid cells (g) in BM, as well as the indel frequencies determined by TIDE analysis (h). i, Multilineage reconstitution analysis of BM collected from mice engrafted with control or edited CD34⁺ HSPCs (from donor β^Aβ^A_#4). Error bars indicate standard deviation. Data are plotted as mean ± SD for (b, f, i). Median of each group with 1 to 3 mice is shown as line for the other panels.