Figure 7.

HSPCs with CRISPR/Cas9-Repaired Rag2 Restore B and T Cell Development In Vivo

(A) Experimental scheme of CRISPR/Cas9-mediated Rag2 correction in HSPCs from Rag2^−/− mice. The targeted HSPCs were then transplanted into irradiated Rag2^−/−cγ^−/− mice, which were analyzed 4 and 8 weeks after reconstitution. HSPCs treated with RNPs only were transplanted as controls. HR efficiency was verified in the edited HSPCs before transplantation using the PCR strategy indicated in Figure 6A.

(B) Percentages of CD19⁺ B and CD3⁺ T cells in the blood of representative control recipients (n = 4) and experimental recipients (n = 5, termed repair), as determined by flow cytometry (left), and summary of the data (right; dots represent individual mice).

(C) FACS analysis showing the percentages of B and T cells in the bone marrow and spleen of the recipients 8 weeks after reconstitution.

(D) Numbers of B and T cells in the bone marrow (black) and spleen (orange) of control and repair groups 8 weeks after transplantation.

(E) Sequencing data showing frequencies of HR (WT) and NHEJ events in B cells from a recipient mouse 8 weeks after reconstitution. The sequences of the NHEJ events reveal diversity (right).