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. 2021 May 13;137(19):2598–2608. doi: 10.1182/blood.2020008503

Figure 4.

Figure 4.

Highly efficient correction of engrafting X-CGD HSPCs by SpCas9/sg mRNA/ODN donor with i53. (A) Peripheral blood (PB) from mice transplanted with X-CGD HSPCs GE with ODN or AAV (CYBB E7-13pA) analyzed at weeks 8, 12, and 26 for engraftment (hCD45+). (B) Percentages of gp91phox+ cells in hCD45+ PB. (C) FACS showing human CD45+ engraftment in mice bone marrow (BM) 18 to 26 weeks after transplant with ODN mutation-repaired X-CGD HSPCs (n = 3 patients). (D) Gp91phox expression in myeloid-differentiated BM human CD45+ cells from NSG transplanted with ODN mutation–repaired X-CGD HSPCs (3 patients). (E) DHR assay showing ROS production in the myeloid-differentiated cells in panel D. (F) Lineage (CD33+ and CD19+) composition in BM, PB, and spleen of transplanted mice. (G) Gp91phox expression in CD33+ myeloid cells in myeloid-differentiated BM and PB. (H) Percentages of gp91phox+ before (in vitro) and after transplant (BM) with ODN-treated cells using Cas9 RNA (±i53) vs RNP (n = 4 experiments; 3 X-CGD patients, 11 mice). (I) Correction rates (% corrected alleles) before (input, 5 days post-electroporation, blue) vs after transplant (red) (n = 3 X-CGD patients). **P < .001.