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Cloning and recombinant expression of mouse coagulation factor X
H H Heidtmann1, R E Kontermann
1Zentrum für Innere Medizin, Abt. Hämatologie und Onkologie, Philipps-Universität Marburg, Germany.
Thrombosis Research
|October 23, 1998
Summary
Researchers developed recombinant mouse coagulation factor X (FX) for therapeutic applications. This engineered mouse FX functions within the human coagulation system, enabling its study in a relevant animal model.
Area of Science:
- Biochemistry
- Molecular Biology
- Hematology
Background:
- Developing novel therapeutic molecules targeting coagulation factor X (FX) requires effective animal models.
- Tumor-associated proteinases offer a potential activation pathway for engineered FX variants.
Purpose of the Study:
- To isolate and characterize mouse coagulation factor X (FX) for use in preclinical studies.
- To establish a functional recombinant mouse FX suitable for evaluating therapeutic variants in vivo.
Main Methods:
- Isolation of mouse coagulation factor X (FX) coding sequence via PCR from liver cDNA.
- Expression of recombinant mouse FX in human embryonic kidney cells.
- Activation of recombinant mouse FX using Russell's viper venom and functional assessment in human FX-deficient plasma.
Main Results:
- A 481-amino acid prepro protein sequence homologous to known FX was deduced.
- Recombinant mouse FX was secreted as a zymogen and could be activated to FXa.
- Purified recombinant mouse FX functionally reconstituted coagulation in human FX-deficient plasma.
Conclusions:
- Recombinant mouse coagulation factor X (FX) is functionally active and interacts with the human coagulation system.
- This engineered mouse FX provides a valuable tool for analyzing therapeutic FX variants in a mouse model.
- Enables the development of novel FX-based therapeutics for various medical conditions.