A single nucleotide mutation in the mouse renin promoter disrupts blood pressure regulation
J. Clin. Invest. Keiji Tanimoto, et al. 118:1006 doi:10.1172/JCI33824 [Go to this article.]

Figure 1
Tg coplacement strategy. (A) Mouse Ren-1^C BAC. The 155-kbp BstI restriction enzyme fragments with 66 kbp and 80 kbp of 5′ and 3′ flanking sequences, respectively, were used for microinjections. Shown are cis-regulatory elements and 9 exons of the gene. (B, upper left) Map of the Ren-1^C promoter region and structure of the targeting vector (pTmRn/mutCNRE), in which WT (open box) and mut CNRE (mut; filled box) sequences as well as homologous sequences to the target locus (thick lines) are included. Restriction enzyme sites with their positions relative to the transcriptional start site (+1) are shown. The loxP2272 and loxP5171 sequences are shown as open and filled triangles, respectively. The targeted locus was generated in E. coli and used to establish TgM lines. Following intercross with Cre-expressing TgM, selective excision in utero of the DNA segment between a pair of loxP5171 or loxP2272 sites generated either WT or mut loci, respectively. (B, upper right) Introduction of the FRT (shaded ovals) and SfiI sites into 3′-UTR of the gene. The targeting and flippase/FRT (FLP/FRT) recombination was carried out in the E. coli strain EL250 (26). The PCR primer set that simultaneously amplifies Tg and endogenous renin transcripts is shown by arrows. (B, bottom) Comparison of WT and mut CNRE promoter sequences. (C, top) Targeting vector (pTmRn/mutRP-2) for modification of the RP-2. Both WT (open circle) and mut (filled circle) RP-2 sequences as well as homologous sequences to the target (thick lines) are included. Tg coplacement was performed as described in B. (C, bottom) Comparison of the WT and mut RP-2 sequences. The HOX/PBX motif is boxed.