Figure 3. Systemic Injection of AAV-Cre Transduces Functional Hematopoietic Stem Cells.

(A) Experimental design. AAVs encoding Cre recombinase were injected into mdx;Ai9 mice carrying the Rosa26-LSL-tdTomato reporter. i.v., intravenous; LSL, LoxP-STOP-LoxP; HSPCs, hematopoietic stem and progenitor cells.

(B) HSC gating strategy (Lin⁻Sca-1⁺c-Kit⁺CD48⁻CD150⁺) from bone marrow (BM) and representative flow cytometric analysis of tdTomato expression within HSCs. LSK: Lin⁻Sca-1⁺c-Kit⁺.

(C) Frequency of HSCs expressing tdTomato. Data points from individual mice are overlaid with mean ± SD. n = 4 mice injected with each AAV serotype per group, n = 3 vehicle-injected mice per group.

(D) Percentage of donor chimerism among live peripheral blood cells of primary recipients at monthly intervals post-transplantation. Data points represent mean ± SD. n = 8 primary recipients per group.

(E) Frequency of primary recipients with tdTomato⁺ multilineage engraftment within the indicated peripheral blood lineages at 16 weeks post-transplant. For each lineage, tdTomato⁺ engraftment was scored based on satisfaction of two criteria: >1% CD45.2⁺ cells and >1% tdTomato⁺ among CD45.2⁺ cells.

(F–I) Frequency of tdTomato⁺ cells among live CD45.2⁺ peripheral blood (F) T cells, (G) B cells, (H) monocytes, and (I) neutrophils in primary recipients at 16 weeks post-transplantation. For each lineage, only recipients exhibiting >1% CD45.2⁺ cells within that lineage are shown. Individual data points are shown overlaid with mean ± SD. n = 6–8 primary recipients per group.

(J) Percentage of tdTomato⁺ HSCs among CD45.2+ HSCs in primary recipients at 6 months post-transplantation Individual data points are shown overlaid with mean ± SD. n = 6–8 primary recipients per group.

vg, viral genomes; Anc80, Anc80L65.

See also Figures S3–S6 and Tables S1 and S2.