Figure 3.

Long-term engraftment of NP-RNP-edited human CD34⁺ cells

(A–C) Schematic of the experimental design. Longitudinal flow cytometric assessment of (B) human chimerism and (C) CD14⁺ monocytes in the PB. (D) PCR validation of CD33 editing in human cells in the PB at weeks (W) 8 and 10 post-transplant. (E) Quantification of PCR bands in (D). (F) Longitudinal tracking of human CD14⁺ monocytes in the PB lacking CD33 exon 2 on the cell surface (CD33^ΔE2) via flow cytometry. (G) Frequency of human chimerism in the bone marrow (BM), spleen, and thymus. (H) Frequency of engrafted human CD34⁺ cells (left y axis) and the HSC-enriched CD34⁺CD90⁺ subset (right y axis) in the BM. (I) Genomic analysis of the CD33 genotype (+/+, wild type; +/−, heterozygous knockout; −/−, homozygous knockout) of individual CD34⁺- and CD34⁺CD90⁺-derived colonies (representative gel pictures shown in Figure S3I). Statistics: (G and H) mean ± SEM, Wilcoxon signed-rank test.