An inducible tissue-specific knockout (KO) technique has been used to study the role of genes in the adult heart. This KO technique circumvents the developmental effect that could otherwise be observed in a tissue-specific KO. The peroxisome proliferator-activated receptor (PPAR) γ is a transcription factor that when activated has been shown to improve vascular remodeling and endothelial function in hypertensive rodents. Here we describe an inducible tissue specific KO protocol used to study the role of PPARγ in smooth muscle cells (SMC) in angiotensin (Ang) II-induced hypertension in adult mice. Inducible VSMC Pparγ KO mice are generated by crossing mice expressing a fusion protein of Cre recombinase with the modified estrogen receptor ligand binding domain (CreER) under the control of the smooth muscle myosin heavy chain (smmhc, myh11) with mice having loxP sites flanking exon 2 of the Pparγ gene (Pparγ ). The SMC Pparγ KO is induced by treating smMHC-CreER/Pparγ mice with the estrogen receptor antagonist tamoxifen causing recombination of the two loxP site by CreER. SMC KO is confirmed by determining mRNA Pparγ levels in aortic media. Presence of the loxP sites is determined by sequencing genomic DNA. Tissue specific expression is assayed using smMHC-CreER/reporter crossed mice.
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