RT Journal Article
T1 Development of a novel and viable knock-in factor V deficiency murine model: Utility for an ultra-rare disease
A1 De Pablo Moreno, Juan Andrés
A1 González-Brusi, Leopoldo
A1 Miguel Batuecas, Andrea
A1 Bermejo Álvarez, Pablo
A1 Revuelta Rueda, Luis
A1 Liras Martín, Antonio
AB Factor V deficiency is a congenital coagulation disorder characterized by the absence or malfunction of factor V (FV). The purpose of this study was to develop a viable FV-deficient mouse model using CRISPR/Cas9 technology. A viable pathological model of the disease was not available to develop new therapies. A previous in silico study was performed to select a mutation causing a mild disease phenotype in humans (Thr1898Met missense). Such mutation was replicated in mice by CRISPR-mediated homology directed repair. Following crossing, homozygous individuals were subjected to coagulometry assays, including FV levels, prothrombin time (PT), and activated partial thromboplastin time (aPTT). The in silico study suggested that the mutation destabilizes FV structure of both mouse and human variants, putatively producing a mild phenotype of the disease in mice. Mendelian inheritance was observed in the offspring. No spontaneous signs of blood clotting disturbances, premature deaths or gestational dysfunctions were observed. FV levels in homozygous animals were 24.5% ± 5.1; 39.7 sec ± 2.8; PT was 61.8% ± 6.3; 23.4 sec ± 1.6 (INR = 1.47 ± 0.12); and aPTT was 46.9 sec ± 3.2. A viable FV-deficient mouse model was generated by introducing a missense mutation in FV. The model exhibits a mild phenotype of the disease, akin to that observed in humans.
PB Public Library of Science (PLOS)
SN 1932-6203
YR 2025
FD 2025-06-02
LK https://hdl.handle.net/20.500.14352/123078
UL https://hdl.handle.net/20.500.14352/123078
LA eng
NO De Pablo-Moreno, J. A., González-Brusi, L., Miguel-Batuecas, A., Bermejo-Álvarez, P., Revuelta, L., & Liras, A. (2025). Development of a novel and viable knock-in factor V deficiency murine model: Utility for an ultra-rare disease. PLOS One, 20(6), e0321864. https://doi.org/10.1371/journal.pone.0321864
NO Funding: AMB received a predoctoral contract from the Community of Madrid, grant number CT85/23; AL received funding for the development of this research from the Association for Research and Cure of Factor V deficiency (ASDEFAV), grant number ASDEFAV/2021-24 (https://unaesperanzaparacelia.org/); JADPM received a predoctoral contract from the Complutense University of Madrid and Banco Santander, grant number CT63/19-CT64/19 (https://www.ucm.es/ct63-19-ct64-19), and PBA received funding for the development of this research from the Spanish Ministry of Science and Innovation, grant number PID2020-117501RB-I00 (https://www.ciencia.gob.es/en/). The sponsors or funders did not play any role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
NO Comunidad de Madrid
NO Association for Research and Cure of Factor V deficiency (ASDEFAV)
NO Universidad Complutense de Madrid
NO Banco Santander
NO Ministerio de Ciencia e Innovación (España)
DS Docta Complutense
RD 24 dic 2025