title: Targeted deletion of the cytosolic domain of tissue factor in mice does not affect development.
abstract: 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.
authors.affiliation: Center for Transgene Technology and Gene Therapy, Flanders Interuniversity Institute for Biotechnology (VIB), KU Leuven, Leuven, B-3000, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: Melis
authors.forename: L
authors.initials: L
authors.lastname: Moons
authors.forename: M
authors.initials: M
authors.lastname: De Mol
authors.forename: J M
authors.initials: JM
authors.lastname: Herbert
authors.forename: N
authors.initials: N
authors.lastname: Mackman
authors.forename: D
authors.initials: D
authors.lastname: Collen
authors.forename: P
authors.initials: P
authors.lastname: Carmeliet
authors.forename: M
authors.initials: M
authors.lastname: Dewerchin
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000098:blood supply; D005314:Embryonic and Fetal Development; D015942:Factor VIIa / Q000494:pharmacology; D015951:Factor Xa / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D017353:Gene Deletion; D018390:Gene Targeting; D006487:Hemostasis; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D018919:Neovascularization, Physiologic; D017434:Protein Structure, Tertiary; D013925:Thromboplastin / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D013927:Thrombosis / Q000209:etiology
chemical_list: D013925:Thromboplastin; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D015942:Factor VIIa; D015951:Factor Xa