Reduced hematopoietic differentiation potential of GATA2-haploinsufficient isogenic iPSC. (A) Exon 6 of the GATA2 gene was targeted to generate knockout isogenic cell lines. (B) Structure of the wild-type GATA2/DNA complex modeled from the corresponding GATA3 structure: sequence RNA targeted for deletion by CRISPR (yellow); the 38-aa long C terminus sequence (red) is truncated and modified because of a premature stop codon; the resulting 13-aa segment has unknown, albeit predictable, conformation in solution; DNA molecule (blue), Zn ions (white), and unmodified GATA2 sequence (green). (C) Isogenic iPSC lines were differentiated into hematopoietic lineage as described previously. Fraction of CD34⁺CD45⁺ cells and the number and type of CFU were examined from independent GATA2 knockout isogenic lines and the 2 corrected patient-specific iPSC lines shown. A minimum of 2 independent experiments was performed. Fraction of CD34⁺CD45⁺ cells was grouped per cell line and GATA2 status. (Left: n = 4, 4, 4, 4, 4, 4, 4, 4, 3, 2, and 2, respectively; right: n = 8, 8, 8, 5, and 5, respectively). (D) Total number of colonies from 1000 sorted CD34⁺CD45⁺ cells per cell line and GATA2 status. (Left: n = 4, 4, 2, 2, 2, 2, 4, 4, 4, 2, and 2, respectively; right: n = 6, 6, 4, 6, and 6, respectively). (E) Fraction of CFU subtype per total number of colonies, respective of mutation types (GATA2^+/+, n = 10; GATA2^+/−, n = 10). Paired Student t test was performed to compare the effect of GATA2 gene status with its isogenic cell line. *P < .05; **P < .01. Nonsignificant P values are not shown.