Figure 1.

runx1^−un mutants survive to adult and recover multilineage hematopoiesis. (A) Schematic representation of the runx1 gene showing the regions (orange bars) targeted with engineered TALENs that generated the runx1^del8, runx1^del25 mutant lines and the CRISPR targets, T1 and T2 (blue bars), used to generate the runx1^del(e3-8) mutant line as shown in the lower panel. The cyan box in the lower panel indicates a 66-bp insertion between the 2 cut sites. Percentage of gata1:dsRed⁺ (B) and cd41:GFP^low or cd41:GFP^high (C) in Tg(gata1:dsRed; cd41:GFP) runx1^del8/del8 runx1^+/del8 controls at 10 and 17 dpf analyzed by flow cytometry (^##P < .01, ^####P < .0001). (D) Kidney histology and blood smear of a representative 22 dpf runx1^del8/del8 appear similar to a runx1^+/del8 control sibling. (E) Stacked bar chart showing the percentage of adult runx1^del/del8, runx1^del2/del25, and runx1^{del(e3-8)/del(e3-8)} recovered from inbred heterozygous parents; red dashed lines indicate the expected Mendelian ratio. (F) Volcano plot showing the differentially expressed genes in the kidneys between adult runx1^−un and wild-type zebrafish. The red dots identify known hematopoietic markers for thrombocytes, myeloid cells, and HSCs that are differentially expressed at FC > 2 and P_adj < .05. (F′) Gene ontology enrichment analysis results for panel F.