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Melis, E, Moons, L, De Mol, M, Herbert, JM, Mackman, N, Collen, D, Carmeliet, P and Dewerchin, M (2001) Targeted deletion of the cytosolic domain of tissue factor in mice does not affect development. Biochem. Biophys. Res. Commun. 286:580-6
The role of the cytosolic domain of tissue factor (TF) in signal transduction and gene regulation was studied in mice with a targeted deletion of the 18 carboxy-terminal intracellular amino acids. This deletion was introduced in exon 6 along with a floxed neo(R) selection cassette in intron 5 using homologous recombination in embryonic stem cells. Removal of the floxed neo(R) cassette by in vivo Cre-mediated loxP recombination yielded TF(+/deltaCT) and TF(deltaCT/deltaCT) mice. In contrast to TF(-/-) mice, TF(+/deltaCT) and TF(deltaCT/deltaCT) mice displayed normal embryonic development, survival, fertility, and blood coagulation. Factor VIIa or factor Xa stimulation produced similar p44/42 MAPK activation in TF(+/+) and TF(deltaCT/deltaCT) fibroblasts. These data, based on expression of a TF(deltaCT) molecule from the endogenous TF locus, provide conclusive proof that the cytosolic domain of TF is not essential for signal transduction in embryogenesis and in physiological postnatal processes.
Animals; Cells, Cultured; Embryo, Mammalian/blood supply; Embryonic and Fetal Development; Factor VIIa/pharmacology; Factor Xa/pharmacology; Fibroblasts/drug effects; Fibroblasts/metabolism; Gene Deletion; Gene Targeting; Hemostasis; Mice; Mitogen-Activated Protein Kinase 1/metabolism; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases/metabolism; Neovascularization, Physiologic; Protein Structure, Tertiary; Thromboplastin/chemistry; Thromboplastin/genetics; Thromboplastin/physiology; Thrombosis/etiology
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