Description

Background:

• F2 encodes the prothrombin protein, which is best known for its role in thrombosis and hemostasis, as well as in cell proliferation, tissue repair, angiogenesis, and maintenance of vascular integrity. This protein is tissue-specific, expressed by the liver and secreted into the plasma.

Targeting strategy:

• The exons 1-14 of mouse F2 gene that encode the whole molecule (ATG to STOP codon) were replaced by human counterparts in B-hF2 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human F2 expression is driven by endogenous mouse F2 promoter, while mouse F2 gene transcription and translation will be disrupted.

Verification: 

• Mouse F2 mRNA were detectable in wild-type C57BL/6JNifdc. Human F2 mRNA was only detectable in homozygous B-hF2 mice.
• Human thrombin was exclusively detectable in homozygous B-hF2 mice.

Application:

• This product is used for pharmacodynamics and safety evaluation of stroke, thrombosis, hemostatic disorders and other blood disorders.

Targeting strategy

Gene targeting strategy for B-hF2 mice. The exons 1-14 of mouse F2 gene that encode the whole molecule (ATG to STOP codon) were replaced by human counterparts in B-hF2 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human F2 expression is driven by endogenous mouse F2 promoter, while mouse F2 gene transcription and translation will be disrupted.

mRNA expression analysis

Strain specific analysis of F2 mRNA expression in wild-type C57BL/6JNifdc mice and B-hF2 mice by RT-PCR. Liver RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hF2 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with human F2 primers. Mouse F2 mRNA were detectable in wild-type C57BL/6JNifdc. Human F2 mRNA was only detectable in homozygous B-hF2 mice.

Protein expression analysis in plasma

Strain specific thrombin expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hF2 mice by ELISA. Plasma was collected from wild-type C57BL/6JNifdc mice (+/+) (male, n=3, 6-week-old) and homozygous B-hF2 mice (H/H) (male, n=3, 6-week-old). Expression level of human thrombin were analyzed by ELISA (Abcam, ab270210). Human thrombin was exclusively detectable in homozygous B-hF2 mice. Values are expressed as mean ± SEM.