Description

Strain Information

Former Names B6;129S4-F8tm1Kaz/J    (Changed: 01-AUG-12 ) Type Mutant Stock; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Mating System Homozygote x Hemizygote         (Female x Male)   01-MAR-06 Species laboratory mouse Generation F?+11N1F5 (03-NOV-08) Generation Definitions   Donating Investigator Haig H. Kazazian, Jr.,   University of Pennsylvania

Description
Mice that are homozygous for the targeted, X chromosome-linked mutant allele are viable and fertile. Homozygous females and carrier males have less than 1% of normal factor VIII activity and exhibit prolonged clotting times. Care should be used when obtaining tail clippings for the purpose of genotyping. Clipped tails of affected mice must be cauterized immediately or the mouse will succumb to excessive blood loss within several hours. Only mice that have reached the age of 4 weeks or older should have their tails clipped. Published reports indicate that spontaneous bleeding into joints or soft tissues, and that no bleeding difficulties are apparent during birth. These mice recapitulate key features of hemophilia A and provide an excellent model for use in exploring gene therapy strategies.

NOTE: It has been the experience of The Jackson Laboratory, in contrast to the primary publication, and possibly due to genetic background differences as a result of continuous inbreeding, animals homozygous or hemizygous for the F8^tm1Kaz allele do exhibit bleeding into joints and soft tissues.

Development
A targeting vector containing a neo cassette was used to disrupt exon 16. The construct was introduced into 129S4/SvJae-derived J1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts.

Control Information

	Control
	101045 B6129SF2/J	(approximate)
Considerations for Choosing Controls

Related Strains

Strains carrying   F8^tm1Kaz allele

017706

STOCK F8tm1Kaz Tg(Alb-F8*R593C)T4Mcal/J

View Strains carrying   F8^tm1Kaz     (1 strain)

Phenotype

Phenotype Information

View Related Disease (OMIM) Terms

Related Disease (OMIM) Terms provided by MGI

Models with phenotypic similarity to human diseases where etiology is unknown or involving genes where ortholog is unknown.

Hemophilia A; HEMA

View Mammalian Phenotype Terms

Mammalian Phenotype Terms provided by MGI

      assigned by genotype

F8^tm1Kaz/F8^tm1Kaz

        involves: 129S4/SvJae * C57BL/6

• mortality/aging
• increased sensitivity to induced morbidity/mortality
  • 2/3 mice die within 2 hours of tail snipping from excessive bleeding   (MGI Ref ID J:36698)
• reproductive system phenotype
• *normal* reproductive system phenotype
  • females exhibit normal fertility and pregnancy   (MGI Ref ID J:36698)
• homeostasis/metabolism phenotype
• abnormal blood coagulation
  • 2/3 mice die within 2 hours of tail snipping from excessive bleeding   (MGI Ref ID J:36698)
• hematopoietic system phenotype
• *normal* hematopoietic system phenotype
  • males exhibit no spontaneous bleeding, illness or reduction in activity up to 1 year of age   (MGI Ref ID J:36698)

F8^tm1Kaz/Y

        involves: 129S4/SvJae * C57BL/6

• mortality/aging
• increased sensitivity to induced morbidity/mortality
  • 2/3 males die within 2 hours of tail snipping from excessive bleeding   (MGI Ref ID J:36698)
  • mice exhibit a reduced ability to clot blood upon tail cutting, with most animals dying within 2 hours of tail snipping unless cauterized   (MGI Ref ID J:24941)
• homeostasis/metabolism phenotype
• abnormal blood coagulation
  • male hemizygous mice exhibit hemophilia   (MGI Ref ID J:24941)
  • mice exhibit a reduced ability to clot blood upon tail cutting, with most animals dying within 2 hours of tail snipping unless cauterized   (MGI Ref ID J:24941)
  • plasma from mutant mice exhibits a prolonged clotting time compared to controls   (MGI Ref ID J:24941)
  • 2/3 males die within 2 hours of tail snipping from excessive bleeding   (MGI Ref ID J:36698)
• hematopoietic system phenotype
• *normal* hematopoietic system phenotype
  • no spontaneous bleeding is observed into joints or soft tissues up to 12 weeks of age   (MGI Ref ID J:24941)
  • males exhibit no spontaneous bleeding, illness or reduction in activity up to 1 year of age   (MGI Ref ID J:36698)
• immune system phenotype
• abnormal immune tolerance
  • mutants develop anti-factor VIII antibodies following administration of factor VIII   (MGI Ref ID J:178969)

View Research Applications

Research Applications

This mouse can be used to support research in many areas including:

F8^tm1Kaz related

Hematological Research
Clotting Defects

Genes & Alleles

Gene & Allele Information provided by MGI

Allele Symbol		F8tm1Kaz
Allele Name		targeted mutation 1, Haig H Kazazian Jr
Allele Type		Targeted (knock-out)
Common Name(s)		E-16; FVIII KO; HemA; MFVIII-16;
Mutation Made By		Haig Kazazian, Jr.,   University of Pennsylvania
Strain of Origin		129S4/SvJae
ES Cell Line Name		J1
ES Cell Line Strain		129S4/SvJae
Gene Symbol and Name		F8, coagulation factor VIII
Chromosome		X
Gene Common Name(s)		AHF; Cf-8; Cf8; DXS1253E; F8B; F8C; FVIII; Factor VIII; HEMA;
Molecular Note		A neomycin resistance cassette replaced 293 bp of sequence, including 7 bp at the 3' end of exon 16 and 286 bp at the 5' end of intron 16. Two different activity assays indicated that the mutated allele produced protein with levels of activity less than 1% of wild-type levels. [MGI Ref ID J:24941]

Genotyping

Genotyping Information

Genotyping Protocols

F8^tm1Kaz, Separated PCR

Helpful Links

Genotyping resources and troubleshooting

References

References provided by MGI

Selected Reference(s)

Bi L; Lawler AM; Antonarakis SE; High KA; Gearhart JD; Kazazian HH Jr. 1995. Targeted disruption of the mouse factor VIII gene produces a model of haemophilia A [letter] Nat Genet 10(1):119-21. [PubMed: 7647782]  [MGI Ref ID J:24941]

Additional References

Fakharzadeh SS; Zhang Y; Sarkar R; Kazazian HH Jr. 2000. Correction of the coagulation defect in hemophilia A mice through factor VIII expression in skin. Blood 95(9):2799-805. [PubMed: 10779424]  [MGI Ref ID J:61846]

F8^tm1Kaz related

Ai D; Pang W; Li N; Xu M; Jones PD; Yang J; Zhang Y; Chiamvimonvat N; Shyy JY; Hammock BD; Zhu Y. 2009. Soluble epoxide hydrolase plays an essential role in angiotensin II-induced cardiac hypertrophy. Proc Natl Acad Sci U S A 106(2):564-9. [PubMed: 19126686]  [MGI Ref ID J:143868]

Bi L; Sarkar R; Naas T; Lawler AM; Pain J; Shumaker SL; Bedian V; Kazazian HH Jr. 1996. Further characterization of factor VIII-deficient mice created by gene targeting: RNA and protein studies. Blood 88(9):3446-50. [PubMed: 8896409]  [MGI Ref ID J:36698]

Bril WS; van Helden PM; Hausl C; Zuurveld MG; Ahmad RU; Hollestelle MJ; Reitsma PH; Fijnvandraat K; van Lier RA; Schwarz HP; Mertens K; Reipert BM; Voorberg J. 2006. Tolerance to factor VIII in a transgenic mouse expressing human factor VIII cDNA carrying an Arg(593) to Cys substitution. Thromb Haemost 95(2):341-7. [PubMed: 16493498]  [MGI Ref ID J:178969]

Chauhan AK; Kisucka J; Lamb CB; Bergmeier W; Wagner DD. 2007. von Willebrand factor and factor VIII are independently required to form stable occlusive thrombi in injured veins. Blood 109(6):2424-9. [PubMed: 17119108]  [MGI Ref ID J:145354]

Dimitrov JD; Dasgupta S; Navarrete AM; Delignat S; Repesse Y; Meslier Y; Planchais C; Teyssandier M; Motterlini R; Bayry J; Kaveri SV; Lacroix-Desmazes S. 2010. Induction of heme oxygenase-1 in factor VIII-deficient mice reduces the immune response to therapeutic factor VIII. Blood 115(13):2682-5. [PubMed: 19890094]  [MGI Ref ID J:160804]

Dumont JA; Liu T; Low SC; Zhang X; Kamphaus G; Sakorafas P; Fraley C; Drager D; Reidy T; McCue J; Franck HW; Merricks EP; Nichols TC; Bitonti AJ; Pierce GF; Jiang H. 2012. Prolonged activity of a recombinant factor VIII-Fc fusion protein in hemophilia A mice and dogs. Blood 119(13):3024-30. [PubMed: 22246033]  [MGI Ref ID J:182551]

Evans GL; Morgan RA. 1998. Genetic induction of immune tolerance to human clotting factor VIII in a mouse model for hemophilia A. Proc Natl Acad Sci U S A 95(10):5734-9. [PubMed: 9576953]  [MGI Ref ID J:47661]

Fakharzadeh SS; Zhang Y; Sarkar R; Kazazian HH Jr. 2000. Correction of the coagulation defect in hemophilia A mice through factor VIII expression in skin. Blood 95(9):2799-805. [PubMed: 10779424]  [MGI Ref ID J:61846]

Follenzi A; Raut S; Merlin S; Sarkar R; Gupta S. 2012. Role of bone marrow transplantation for correcting hemophilia A in mice. Blood 119(23):5532-42. [PubMed: 22368271]  [MGI Ref ID J:186576]

Greene TK; Wang C; Hirsch JD; Zhai L; Gewirtz J; Thornton MA; Miao HZ; Pipe SW; Kaufman RJ; Camire RM; Arruda VR; Kowalska MA; Poncz M. 2010. In vivo efficacy of platelet-delivered, high specific activity factor VIII variants. Blood 116(26):6114-22. [PubMed: 20852129]  [MGI Ref ID J:167399]

Hakobyan N; Kazarian T; Jabbar AA; Jabbar KJ; Valentino LA. 2004. Pathobiology of hemophilic synovitis I: overexpression of mdm2 oncogene. Blood 104(7):2060-4. [PubMed: 15172967]  [MGI Ref ID J:93755]

Hrachovinova I; Cambien B; Hafezi-Moghadam A; Kappelmayer J; Camphausen RT; Widom A; Xia L; Kazazian HH Jr; Schaub RG; McEver RP; Wagner DD. 2003. Interaction of P-selectin and PSGL-1 generates microparticles that correct hemostasis in a mouse model of hemophilia A. Nat Med 9(8):1020-5. [PubMed: 12858167]  [MGI Ref ID J:99599]

Kawasaki T; Dewerchin M; Lijnen HR; Vreys I; Vermylen J; Hoylaerts MF. 2001. Mouse carotid artery ligation induces platelet-leukocyte-dependent luminal fibrin, required for neointima development. Circ Res 88(2):159-66. [PubMed: 11157667]  [MGI Ref ID J:115383]

Litzinger MT; Su Y; Lei TC; Soukhareva N; Scott DW. 2005. Mechanisms of gene therapy for tolerance: B7 signaling is required for peptide-IgG gene-transferred tolerance induction. J Immunol 175(2):780-7. [PubMed: 16002674]  [MGI Ref ID J:100709]

Maroney SA; Cooley BC; Ferrel JP; Bonesho CE; Nielsen LV; Johansen PB; Hermit MB; Petersen LC; Mast AE. 2012. Absence of hematopoietic tissue factor pathway inhibitor mitigates bleeding in mice with hemophilia. Proc Natl Acad Sci U S A 109(10):3927-31. [PubMed: 22355108]  [MGI Ref ID J:182144]

Matsui H; Shibata M; Brown B; Labelle A; Hegadorn C; Andrews C; Chuah M; VandenDriessche T; Miao CH; Hough C; Lillicrap D. 2009. A murine model for induction of long-term immunologic tolerance to factor VIII does not require persistent detectable levels of plasma factor VIII and involves contributions from Foxp3+ T regulatory cells. Blood 114(3):677-85. [PubMed: 19458355]  [MGI Ref ID J:150739]

Meeks SL; Cox CL; Healey JF; Parker ET; Doshi BS; Gangadharan B; Barrow RT; Lollar P. 2012. A major determinant of the immunogenicity of factor VIII in a murine model is independent of its procoagulant function. Blood 120(12):2512-20. [PubMed: 22855607]  [MGI Ref ID J:191310]

Mei B; Pan C; Jiang H; Tjandra H; Strauss J; Chen Y; Liu T; Zhang X; Severs J; Newgren J; Chen J; Gu JM; Subramanyam B; Fournel MA; Pierce GF; Murphy JE. 2010. Rational design of a fully active, long-acting PEGylated factor VIII for hemophilia A treatment. Blood 116(2):270-9. [PubMed: 20194895]  [MGI Ref ID J:162833]

Miao CH; Harmeling BR; Ziegler SF; Yen BC; Torgerson T; Chen L; Yau RJ; Peng B; Thompson AR; Ochs HD; Rawlings DJ. 2009. CD4+FOXP3+ regulatory T cells confer long-term regulation of factor VIII-specific immune responses in plasmid-mediated gene therapy-treated hemophilia mice. Blood 114(19):4034-44. [PubMed: 19713458]  [MGI Ref ID J:154194]

Milanov P; Ivanciu L; Abriss D; Quade-Lyssy P; Miesbach W; Alesci S; Tonn T; Grez M; Seifried E; Schuttrumpf J. 2012. Engineered factor IX variants bypass FVIII and correct hemophilia A phenotype in mice. Blood 119(2):602-11. [PubMed: 22031860]  [MGI Ref ID J:181577]

Neyman M; Gewirtz J; Poncz M. 2008. Analysis of the spatial and temporal characteristics of platelet-delivered factor VIII-based clots. Blood 112(4):1101-8. [PubMed: 18559671]  [MGI Ref ID J:139175]

Ohlfest JR; Frandsen JL; Fritz S; Lobitz PD; Perkinson SG; Clark KJ; Nelsestuen G; Key NS; McIvor RS; Hackett PB; Largaespada DA. 2005. Phenotypic correction and long-term expression of factor VIII in hemophilic mice by immunotolerization and nonviral gene transfer using the Sleeping Beauty transposon system. Blood 105(7):2691-8. [PubMed: 15576475]  [MGI Ref ID J:105059]

Purohit VS; Ramani K; Sarkar R; Kazazian HH Jr; Balasubramanian SV. 2005. Lower inhibitor development in hemophilia A mice following administration of recombinant factor VIII-O-phospho-L-serine complex. J Biol Chem 280(18):17593-600. [PubMed: 15728582]  [MGI Ref ID J:99097]

Qadura M; Waters B; Burnett E; Chegeni R; Bradshaw S; Hough C; Othman M; Lillicrap D. 2009. Recombinant and plasma-derived factor VIII products induce distinct splenic cytokine microenvironments in hemophilia A mice. Blood 114(4):871-80. [PubMed: 19411636]  [MGI Ref ID J:150765]

Qian J; Collins M; Sharpe AH; Hoyer LW. 2000. Prevention and treatment of factor VIII inhibitors in murine hemophilia A. Blood 95(4):1324-9. [PubMed: 10666206]  [MGI Ref ID J:110275]

Ragni MV; Wu W; Liang X; Hsieh CC; Cortese-Hassett A; Lu L. 2009. Factor VIII-pulsed dendritic cells reduce anti-factor VIII antibody formation in the hemophilia A mouse model. Exp Hematol 37(6):744-54. [PubMed: 19463774]  [MGI Ref ID J:151239]

Rastegarlari G; Pegon JN; Casari C; Odouard S; Navarrete AM; Saint-Lu N; van Vlijmen BJ; Legendre P; Christophe OD; Denis CV; Lenting PJ. 2012. Macrophage LRP1 contributes to the clearance of von Willebrand factor. Blood 119(9):2126-34. [PubMed: 22234691]  [MGI Ref ID J:182558]

Rossi G; Sarkar J; Scandella D. 2001. Long-term induction of immune tolerance after blockade of CD40-CD40L interaction in a mouse model of hemophilia A. Blood 97(9):2750-7. [PubMed: 11313267]  [MGI Ref ID J:70448]

Sack BK; Merchant S; Markusic DM; Nathwani AC; Davidoff AM; Byrne BJ; Herzog RW. 2012. Transient B cell depletion or improved transgene expression by codon optimization promote tolerance to factor VIII in gene therapy. PLoS One 7(5):e37671. [PubMed: 22655063]  [MGI Ref ID J:187309]

Sen D; Chapla A; Walter N; Daniel V; Srivastava A; Jayandharan GR. 2013. Nuclear factor (NF)-kappaB and its associated pathways are major molecular regulators of blood-induced joint damage in a murine model of hemophilia. J Thromb Haemost 11(2):293-306. [PubMed: 23231432]  [MGI Ref ID J:195442]

Skupsky J; Zhang AH; Su Y; Scott DW. 2009. A role for thrombin in the initiation of the immune response to therapeutic factor VIII. Blood 114(21):4741-8. [PubMed: 19794141]  [MGI Ref ID J:155485]

Stennicke HR; Kjalke M; Karpf DM; Balling KW; Johansen PB; Elm T; Ovlisen K; Moller F; Holmberg HL; Gudme CN; Persson E; Hilden I; Pelzer H; Rahbek-Nielsen H; Jespersgaard C; Bogsnes A; Pedersen AA; Kristensen AK; Peschke B; Kappers W; Rode F; Thim L; Tranholm M; Ezban M; Olsen EH; Bjorn SE. 2013. A novel B-domain O-glycoPEGylated FVIII (N8-GP) demonstrates full efficacy and prolonged effect in hemophilic mice models. Blood 121(11):2108-16. [PubMed: 23335368]  [MGI Ref ID J:195866]

Waters B; Qadura M; Burnett E; Chegeni R; Labelle A; Thompson P; Hough C; Lillicrap D. 2009. Anti-CD3 prevents factor VIII inhibitor development in hemophilia A mice by a regulatory CD4+CD25+-dependent mechanism and by shifting cytokine production to favor a Th1 response. Blood 113(1):193-203. [PubMed: 18815284]  [MGI Ref ID J:144256]

Xu D; Alipio Z; Fink LM; Adcock DM; Yang J; Ward DC; Ma Y. 2009. Phenotypic correction of murine hemophilia A using an iPS cell-based therapy. Proc Natl Acad Sci U S A 106(3):808-13. [PubMed: 19139414]  [MGI Ref ID J:143861]

Yarovoi H; Nurden AT; Montgomery RR; Nurden P; Poncz M. 2005. Intracellular interaction of von Willebrand factor and factor VIII depends on cellular context: lessons from platelet-expressed factor VIII. Blood 105(12):4674-6. [PubMed: 15731176]  [MGI Ref ID J:107451]

Zhang AH; Skupsky J; Scott DW. 2011. Effect of B-cell depletion using anti-CD20 therapy on inhibitory antibody formation to human FVIII in hemophilia A mice. Blood 117(7):2223-6. [PubMed: 21160036]  [MGI Ref ID J:169609]

Health & husbandry

Health & Colony Maintenance Information

Animal Health Reports

Room Number           AX12

Colony Maintenance

Breeding & Husbandry	This strain originated on a B6;129S4 background. It is maintained by crossing homozygous females x hemizygous males on the same background. Expected coat color: Agouti and black. Homozygous females and carrier males have less than 1% of normal factor VIII activity and exhibit prolonged clotting times. Care should be exercise when obtaining tail clippings for the purpose of genotyping. Clipped tails of affected mice must be cauterized immediately or the mouse will succumb to excessive blood loss within several hours. NOTE: It has been the experience of The Jackson Laboratory, in contrast to the primary publication, and possibly due to genetic background differences as a result of continuous inbreeding, animals homozygous or hemizygous for the F8tm1Kaz allele do exhibit bleeding into joints and soft tissues.
Mating System	Homozygote x Hemizygote         (Female x Male)   01-MAR-06
Diet Information	LabDiet® 5K52/5K67

Pricing and Purchasing

Pricing, Supply Level & Notes, Controls

General Supply Notes

• Genomic DNA is available for this strain from the Mouse DNA Resource.

Control Information

	Control
	101045 B6129SF2/J	(approximate)
Considerations for Choosing Controls
Control Pricing Information for Genetically Engineered Mutant Strains.

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