Figure 5.

Efficient CRISPR/Cas9 RNP-based ELANE knockout in HSPC. (A) Scheme of the generation of ELANE KO HSPC using electroporation with ELANE-specific CRISPR/Cas9-gRNA ribonucleoprotein (RNP). (B) TIDE results of edited CD34⁺ HSPC at day 7 and 14 of liquid culture differentiation. (C and D) Hematopoietic stem and progenitor cells (HSPC) of healthy controls (C), or two CN patients (D) were electroporated with ELANE-specific CRISPR/Cas9 RNP, on day 14 of culture, cells were lysed in Laemmli buffer and Western blotting (WB) analysis using antineutrophil elastase (NE) antibody against C-terminus of NE was performed, staining with α-tubulin antibody was used as loading control. Representative WB images of cells from two independent experiments are depicted.