Figure 1.

Adamts16 expression and impact of targeted exon deletion. (A–D) Wholemount in situ hybridisation (WMISH) with Adamts16 probe reveals expression at all developmental stages between 11.5 and 14.5 dpc in XY gonads. Expression is prominent in the testis cords (B–D); (E–H) Significant expression in XX gonads is detected only at 14.5 dpc (H); (I) Exon 5 of Adamts16, encoding part of the peptidase domain, was deleted using CRISPR/Cas9-mediated genome editing (not to scale). See Materials and Methods; (J) WMISH reveals Adamts16 in control (+/+) gonad at 12.5 dpc, but negligible expression in the homozygous mutant (−/−) gonad at the same stage; signal in the mutant gonad was visible upon microscopic examination but was insufficient for photographic reproduction. (K) qRT-PCR shows dramatic reduction in Adamts16 expression in homozygous mutant kidneys (−/−), and intermediate levels in heterozygotes (+/−). In all images in (A–H), the developing gonad is to the left of the white dotted line and the mesonephros is to the right. ***p ≤ 0.01 (student’s t-test).