Model validation analysis
The experimental animals were randomly divided into groups and given NS(normal saline) or 1 mg/kg NovoSeven by tail vein injection, 30 minutes after injection of the drug, blood was collected from the abdominal aorta to detect the blood coagulation index: activated partial thromboplastin time APTT.
The results showed that APTT values in F8 KO mice were much higher than those in WT mice, and APTT returned to normal values after injection of NovoSeven (recombinant human coagulation factor VIIa).
The results showed that B-F8 KO mice can be used as a powerful tool for the validation of anticoagulant efficacy.
Primer Sequence (5’-3’) Tm (℃) Product size
F8-WT-F ACTGTGGCTGGGTGTAGTAGCAC 63 WT: 356 F8-WT-R TTTCTCCAAGTTGCAGGTCAAGGCT 60 F8-WT-F ACTGTGGCTGGGTGTAGTAGCAC 63 Mut: ~400
F8-Mut-R CGCATAATACACCCTGGTCACACTCAC 63
NovoSeven efficacy evaluation
Animals were randomly divided into Control group or NovoSeven (1 mg/kg), After 30 minutes treatment, the abdominal aorta was collected blood for ATPP (activated partial thromboplastin time) detection. The results showed that the APTT recovered to normal after drug treatment compared with the control group.
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