Fig. 3

CRISPR-Cas9-generated Bex3 mutant alleles show skull and brain abnormalities. a Schematic representation of the mouse Bex3 locus (mm10; chrX:136,270,126-136,272,051) depicting exon-intron structure (non-coding and coding exons represented as black and blue rectangles, respectively). Two sgRNAs (red arrows) were used to generate CRISPR-Cas9-mediated deletions. The altered proteins potentially produced by the edited alleles that were selected to generate homozygous mice are shown below. In the Bex3^KO line, the deletion caused a frameshift in the open reading frame (region in gray) leading to the appearance of a premature STOP codon. The deletion in the Bex3^Δ(24–72) line removed 54 amino acids from the central core of the protein. AD, pro-apoptotic domain; CC, coiled coil domain; NES, nuclear export signal. b, c Morphometric analyses of skulls from wild-type and mutant Bex3^KO 6-week-old males employing a total of 24 anatomical measurements b revealed that Bex3 dysfunction led to cranial abnormalities in frontal bone and skull height c (the complete analysis can be found in Additional file 1: Fig. S10). Measurements were normalized to maximum skull length (D0) and expressed relative to controls (black horizontal line). Deviations of 5% with respect to controls are shown as dotted red and green horizontal lines. Results are presented as mean ± SEM (N ≥ 4); *p < 0.05, one-way ANOVA. d, e Bex3 mutant brains showed altered brain morphology as evidenced by gross d and refined e anatomical measurements, concomitant with enlarged ventricular surfaces. Scale bar: 1 mm. Results in d and e are presented as mean ± SEM (N ≥ 5); *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001, one-way ANOVA