Serrano Ramos, Luis JavierGarcía Arranz, MarianoDe Pablo Moreno, Juan AndrésSegovia, José CarlosOlivera Salazar, RocíoGarcía Olmo, DamiánLiras Martín, Antonio2024-11-202024-11-202022Serrano, L. J., Garcia‐arranz, M., De Pablo‐Moreno, J. A., Segovia, J. C., Olivera‐salazar, R., Garcia‐olmo, D., & Liras, A. (2022). Development and Characterization of a Factor V‐Deficient CRISPR Cell Model for the Correction of Mutations. International Journal of Molecular Sciences, 23(10). https://doi.org/10.3390/IJMS231058021661-659610.3390/ijms23105802https://hdl.handle.net/20.500.14352/110863This study was supported by the following public grants: Health Research, Instituto de Salud Carlos III; European Regional Development (RETICS RD 16/0011/00011 and RD 16/0011/0013); Directorate General for Research of the Community of Madrid (AvanCell-CM; Ref. S2017/BMD-3692); State Research Agency (PID2020-119637RB-I00). It also received funds from the Association for the Research and Cure of Factor V Deficiency (ASDEFAV).Factor V deficiency, an ultra-rare congenital coagulopathy, is characterized by bleeding episodes that may be more or less intense as a function of the levels of coagulation factor activity present in plasma. Fresh-frozen plasma, often used to treat patients with factor V deficiency, is a scarcely effective palliative therapy with no specificity to the disease. CRISPR/Cas9-mediated gene editing, following precise deletion by non-homologous end-joining, has proven to be highly effective for modeling on a HepG2 cell line a mutation similar to the one detected in the factor V-deficient patient analyzed in this study, thus simulating the pathological phenotype. Additional CRISPR/Cas9-driven non-homologous end-joining precision deletion steps allowed correction of 41% of the factor V gene mutated cells, giving rise to a newly developed functional protein. Taking into account the plasma concentrations corresponding to the different levels of severity of factor V deficiency, it may be argued that the correction achieved in this study could, in ideal conditions, be sufficient to turn a severe phenotype into a mild or asymptomatic one.engAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/journal article1422-0067https://doi.org/10.3390/ijms23105802https://www.mdpi.com/1422-0067/23/10/5802open access575:61616-056.7612.1577.2Gene therapyGene editingCRISPRFactor V deficiencyCoagulopathiesRare diseasesGenética médicaHematologíaFisiologíaBiología molecular (Biología)3201.02 Genética Clínica3207.08 Hematología2410.10 Fisiología Humana2415 Biología Molecular