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 Plos one
YR 2025
FD 2025-06-02
LK https://hdl.handle.net/20.500.14352/120843
UL https://hdl.handle.net/20.500.14352/120843
LA eng
NO De Pablo-Moreno JA, 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 Author contributions Conceptualization: Juan A. De Pablo-Moreno, Pablo Bermejo-Álvarez, Antonio Liras. Formal analysis: Juan A. De Pablo-Moreno, Leopoldo González-Brusi, Andrea Miguel-Batuecas, Pablo Bermejo-Álvarez, Luis Revuelta, Antonio Liras. Funding acquisition: Pablo Bermejo-Álvarez, Luis Revuelta, Antonio Liras. Investigation: Juan A. De Pablo-Moreno, Leopoldo González-Brusi, Andrea Miguel-Batuecas, Pablo Bermejo-Álvarez, Antonio Liras. Methodology: Juan A. De Pablo-Moreno, Leopoldo González-Brusi, Andrea Miguel-Batuecas, Pablo Bermejo-Álvarez, Luis Revuelta, Antonio Liras. Project administration: Antonio Liras. Resources: Pablo Bermejo-Álvarez, Luis Revuelta, Antonio Liras. Software: Juan A. De Pablo-Moreno, Andrea Miguel-Batuecas. Supervision: Pablo Bermejo-Álvarez, Luis Revuelta, Antonio Liras. Writing – original draft: Juan A. De Pablo-Moreno, Andrea Miguel-Batuecas, Antonio Liras. Writing – review & editing: Juan A. De Pablo-Moreno, Andrea Miguel-Batuecas, Leopoldo González-Brusi, Pablo Bermejo-Álvarez, Luis Revuelta, Antonio Liras
NO Comunidad de Madrid
NO Universidad Complutense de Madrid- Banco Santander
NO Ministerio de Ciencia, Innovación y Universidades ( España)
DS Docta Complutense
RD 19 dic 2025