Sex-specific deficits in neurite density and white matter integrity are associated with targeted disruption of exon 2 of the Disc1 gene in the rat

a The selected strategy was a redundant 2-target CRPSPR/Cas9 approach (low transcript complexity, large targetable N-terminus exon). Two target sites in exon 2 were selected in the second coding exon. b Expected target 1 and 2 cleavage sites are indicated (arrowhead), and target 1 and 2 PAMs (green) are on the sense and antisense strand, respectively. Line #860 carried a 371？bp excision, verified by Sanger sequencing. c PCR of the target region generated a wild-type band for the unedited Sprague-Dawley (SD) sample, two bands for the #860 sample corresponding to the wild-type band and the expected deletion, and no bands for the no-template control (NTC). d Western blot of Disc1 protein from Sprague–Dawley and Disc1 svΔ2 rat brain samples, performed with DSMS/D22 rabbit anti-DISC1 598-785 antibody. From left to right: Sprague-Dawley female rat, Disc1 svΔ2 female rat, Sprague-Dawley male rat, and Disc1 svΔ2 male rat. Western blotting shows absence of an endogenous Disc1 svΔ2 isoform in male and female animals (red arrow) in Disc1 svΔ2 male and female sample