Targeted Deletion of the Cytosolic Domain of Tissue Factor in Mice Does Not Affect Development

• Published in: Biochemical and biophysical research communications Vol. 286; no. 3; pp. 580 - 586
• Main Authors: Melis, Els, Moons, Lieve, De Mol, Maria, Herbert, Jean-Marc, Mackman, Nigel, Collen, Désiré, Carmeliet, Peter, Dewerchin, Mieke
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 24.08.2001
• Subjects: Animals; blood coagulation; Cells, Cultured; cytosolic deletion; Embryo, Mammalian - blood supply; embryogenesis; 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; p42 protein; p44 protein; Protein Structure, Tertiary; signal transduction; TF gene; Thromboplastin - chemistry; Thromboplastin - genetics; Thromboplastin - physiology; Thrombosis - etiology; tissue factor; gene targeting; embryogenesis; tissue factor; cytosolic deletion; signal transduction; blood coagulation
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.2001.5425

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 neoR selection cassette in intron 5 using homologous recombination in embryonic stem cells. Removal of the floxed neoR cassette by in vivo Cre-mediated loxP recombination yielded TF+/δCT and TFδCT/δCT mice. In contrast to TF−/− mice, TF+/δCT and TFδCT/δCT 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 TFδCT/δCT fibroblasts. These data, based on expression of a TFδCT 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.