Targeted deletion of the cytosolic domain of tissue factor in mice does not affect development.
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.
Melis
E
E
Center for Transgene Technology and Gene Therapy, Flanders Interuniversity Institute for Biotechnology (VIB), KU Leuven, Leuven, B-3000, Belgium.
Moons
L
L
De Mol
M
M
Herbert
J M
JM
Mackman
N
N
Collen
D
D
Carmeliet
P
P
Dewerchin
M
M
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
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