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. 2004 Apr 15;18(8):901–911. doi: 10.1101/gad.291004

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Copyright © 2004, Cold Spring Harbor Laboratory Press

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Figure 1. — Knockout of the Hey1 locus and embryonic death of Hey1/2 DKO embryos. (A) The targeting construct is depicted above the mouse genomic Hey1 locus containing five exons. Homology regions of 3 kbp (BglII/Ecl136II) and 1.4 kbp (HindIII) containing mainly promoter and 3′untranslated regions, respectively, were used for recombination. The PGK-neo cassette (Neo) is flanked by loxP sites (triangles) for subsequent removal by cre-recombinase. The HSV-TK (TK) cassette was included for negative selection. Flanking genomic probes used for Southern blot hybridization are highlighted by thick lines. (B) BglII; (E) Ecl136II; (H) HindIII; (*) stop codon; (pBS) pBluescript cloning vector backbone. (B) At E10.5, the Hey1/2 DKO embryos can be clearly identified. They are much smaller compared to a het/het littermate (fourth embryo), and they lack prominent mes- and telencephalic vesicles (*). The extensive pericardial sac is highlighted by arrows. (C,D) Already at E9.5, most DKO embryos suffer from massive hemorrhage in the trunk (arrowheads, C) or the pericardial sac (D).

Figure 1. — Knockout of the Hey1 locus and embryonic death of Hey1/2 DKO embryos. (A) The targeting construct is depicted above the mouse genomic Hey1 locus containing five exons. Homology regions of 3 kbp (BglII/Ecl136II) and 1.4 kbp (HindIII) containing mainly promoter and 3′untranslated regions, respectively, were used for recombination. The PGK-neo cassette (Neo) is flanked by loxP sites (triangles) for subsequent removal by cre-recombinase. The HSV-TK (TK) cassette was included for negative selection. Flanking genomic probes used for Southern blot hybridization are highlighted by thick lines. (B) BglII; (E) Ecl136II; (H) HindIII; (*) stop codon; (pBS) pBluescript cloning vector backbone. (B) At E10.5, the Hey1/2 DKO embryos can be clearly identified. They are much smaller compared to a het/het littermate (fourth embryo), and they lack prominent mes- and telencephalic vesicles (*). The extensive pericardial sac is highlighted by arrows. (C,D) Already at E9.5, most DKO embryos suffer from massive hemorrhage in the trunk (arrowheads, C) or the pericardial sac (D).