Gain-of-function mutant of angiotensin II receptor, type 1A, causes hypertension and cardiovascular fibrosis in mice
J. Clin. Invest. Sandrine Billet, et al. 117:1914 doi:10.1172/JCI28764 [Go to this article.]

Figure 1
Targeted knockin of the Agtr1a gene by homologous recombination into ES cells and mouse. (A) Schematic representation of the targeting vector and partial restriction map of the Agtr1a gene locus before and after homologous recombination. The top line shows the structure of AT_1AWT locus containing exon 3 of the gene with coding sequence (black box) and the 5′ and 3′ untranslated sequences (gray boxes). Exon 3 is flanked by 3 kb of intron 2 (upstream) and 2 kb of noncoding sequence (downstream). The targeting construct containing AT_1AMUT and a hygromycin cassette (hygro) was used to replace the WT BamHI (B) KpnI (K) fragment within AT_1A locus (middle line). The lower line shows the targeted AT_1A locus with the internal probe (probe 1) and 3′ external probe (probe 2) used for Southern blot analysis. (B) Southern blot analysis of targeted ES cell DNA using internal probe 1 with SpeI digestion (left) and using probe 2 with SacI digestion (right). C, control (targeting construct). –/–, WT cells; +/–, heterozygous recombinant cells. (C) Southern blot analysis of tail DNA samples from F₂ offspring (generated by crossing heterozygous mice from the first generation [F₁ mice]) digested with SacI and probed with the 3′ external probe (probe 2). –/–, WT mice; +/–, heterozygous transgenic mice; +/+, homozygous transgenic mice; S, SpeI; B, BamHI; G, BglII; X, XbaI; A, SacI; K, KpnI; N, NotI; E, EcoRI.