Figure 1. Generation of Sox17^GFP/GFP Mice.

(A) Schematic representation of the Sox17 targeted allele. The genomic structure after neo-cassette removal is shown on the fourth line. B and R indicate BamHI and EcoRV digestion sites, respectively.

(B) Genotypes of heterozygous F1 mice (before neo excision) and F2 mice (after neo excision) from two independent lines were confirmed by Southern blot. Tail genomic DNA was digested with EcoRV and BamHI for 5′ and 3′ probe hybridization, respectively.

(C) The lack of Sox17 expression in Sox17^GFP/GFP embryos from both lines of mice was confirmed by RT-PCR using primers that amplified the Sox17 5′ untranslated region (which was expressed from the targeted allele), the coding sequence (not expressed), and the Sox17 3′ untranslated region (not expressed).

(D) At E11.5, Sox17^GFP/GFP embryos were much smaller than Sox17^GFP/+ embryos and exhibited severe defects in posterior patterning (arrow) and body axis rotation (arrowheads).

(E) Genotypes of progeny from Sox17^GFP/+ intercrosses revealed Mendelian inheritance up to E12.5 but loss of Sox17-deficienct embryos by E13.5.