Figure 5.

Engraftment of human genome-edited HSPCs in the CNS of NSG-SGM3 mice

(A) Representative flow cytometry plots showing mouse (mCD45+ CD11b+), human (hCD45+ CD11b+), and edited (YFP+) human microglia-like cells isolated from brain tissue of transplanted NSG-SGM3 mice. (B) Purity of murine microglia (mCD45+ CD11b+) among total myeloid CD45+ cells. (C and D) Characterization of human cells among (C) total myeloid CD45+ cells and (D) human CD45+ cell populations. (E) Proportion of human alleles measured by ddPCR in the microglia preparations, normalized by mouse genomic DNA. (F and G) Characterization of edited YFP+ human HSPCs among (F) total CD45+ and (G) human CD45+ cell populations. (H) Fraction of edited human CCR5 alleles in cells isolated from microglia preparations. Dashed line refers to the fraction of edited alleles in the transplanted HSPC population (input). (I) Representative brain sections of transplanted mice showing YFP expression (natural fluorescence) from edited HSPCs and staining for nuclei (DAPI) and for the specific human marker Ku80. Scale bar is 100 μm. Medians shown; each dot represents a mouse, n = 4.