WHIM allele–inactivated HSPCs have a selective advantage for engraftment to reconstitute hematopoietic cells. (A) Flow cytometry plot of sorted CD11b⁺ cells in blood from a recipient mouse 112 days after transplantation. DNA was isolated for analysis of allele-specific edits from the sorted CD11b⁺ cells, which were derived from RNP-transfected and mock-transfected HSPCs post transplantation, as described in Figure 3. (B) PCR cloning design for sequencing Cxcr4 that covers both the CRISPR/Cas9 PAM site and the WHIM mutation site. (C) Edit analysis for the sorted CD11b⁺ cells shown in panel A. Edits were scored on the wild-type and WHIM alleles for CD11b⁺ cells derived from either the mock-transfected or Cxcr4-sgRNA/Cas9 RNP–transfected donor HSPCs. Eighteen and 25 clones were sequenced for the CD11b⁺ cells derived from mock-transfected and Cxcr4-sgRNA/Cas9 RNP–transfected HSPCs, respectively. (D) Flow cytometry plot of LSK cells sorted from BM of the same recipient mouse shown in panel A 381 days after transplantation. (E) Edit analysis for the sorted LSK cell populations shown in panel D. A total of 6 and 10 clones were sequenced for the cells derived from the mock-transfected and Cxcr4-sgRNA/Cas9 RNP–transfected HSPCs, respectively.