A rat model of severe VWD by elimination of the VWF gene using CRISPR/Cas9.
CRISPR
factor VIII
rat model
severe von Willebrand disease
von Willebrand factor
Journal
Research and practice in thrombosis and haemostasis
ISSN: 2475-0379
Titre abrégé: Res Pract Thromb Haemost
Pays: United States
ID NLM: 101703775
Informations de publication
Date de publication:
Jan 2020
Jan 2020
Historique:
received:
09
09
2019
revised:
02
10
2019
accepted:
06
10
2019
entrez:
29
1
2020
pubmed:
29
1
2020
medline:
29
1
2020
Statut:
epublish
Résumé
Von Willebrand Disease (VWD) is the most common inherited bleeding disorder, caused by quantitative and qualitative changes in von Willebrand factor (VWF). The biology of VWD, studied in canine, porcine, and murine models, differ in species-specific biology of VWF and the amenability to experimental manipulations such as phlebotomy. The factor VIII (FVIII) levels in these models are higher than in humans with type 3 VWD, suggesting functional differences between FVIII and VWF.ObjectivesTo develop a VWF knock out (VWF The entire VWF gene was eliminated in Sprague-Dawley (Crl:SD) rats via CRISPR/Cas9-mediated gene editing. VWF antigen (VWF:Ag), VWF propeptide, and VWF collagen IV binding (VWF:CB4) levels were determined by ELISA assays and FVIII chromogenic activity (FVIII:C) levels by chromogenic FVIII assays. Lateral tail veins were transected to measure bleeding time. VWF Breeding of VWF ± rats yielded VWF This rat model of severe VWD due to elimination of the entire VWF gene recapitulates the severe secondary deficiency of FVIII seen in human type 3 VWD and facilitates the study of VWF and FVIII and their interactions.
Sections du résumé
BACKGROUND
BACKGROUND
Von Willebrand Disease (VWD) is the most common inherited bleeding disorder, caused by quantitative and qualitative changes in von Willebrand factor (VWF). The biology of VWD, studied in canine, porcine, and murine models, differ in species-specific biology of VWF and the amenability to experimental manipulations such as phlebotomy. The factor VIII (FVIII) levels in these models are higher than in humans with type 3 VWD, suggesting functional differences between FVIII and VWF.ObjectivesTo develop a VWF knock out (VWF
METHODS
METHODS
The entire VWF gene was eliminated in Sprague-Dawley (Crl:SD) rats via CRISPR/Cas9-mediated gene editing. VWF antigen (VWF:Ag), VWF propeptide, and VWF collagen IV binding (VWF:CB4) levels were determined by ELISA assays and FVIII chromogenic activity (FVIII:C) levels by chromogenic FVIII assays. Lateral tail veins were transected to measure bleeding time. VWF
RESULTS
RESULTS
Breeding of VWF ± rats yielded VWF
CONCLUSION
CONCLUSIONS
This rat model of severe VWD due to elimination of the entire VWF gene recapitulates the severe secondary deficiency of FVIII seen in human type 3 VWD and facilitates the study of VWF and FVIII and their interactions.
Identifiants
pubmed: 31989086
doi: 10.1002/rth2.12280
pii: S2475-0379(22)01953-7
pmc: PMC6971331
doi:
Types de publication
Journal Article
Langues
eng
Pagination
64-71Subventions
Organisme : NHLBI NIH HHS
ID : R01 HL112614
Pays : United States
Organisme : NHLBI NIH HHS
ID : P01 HL144457
Pays : United States
Organisme : NHLBI NIH HHS
ID : R24 HL114474
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL102035
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL126810
Pays : United States
Organisme : NHLBI NIH HHS
ID : P01 HL081588
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL139847
Pays : United States
Informations de copyright
© 2019 The Authors. Research and Practice in Thrombosis and Haemostasis published by Wiley Periodicals, Inc on behalf of International Society on Thrombosis and Haemostasis.
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