Arranz Solís, DavidRegidor-Cerrillo, JavierLourido, SebastianOrtega Mora, Luis MiguelSaeij, Jeroen2024-01-302024-01-302018Arranz-Solís, D., Regidor-Cerrillo, J., Lourido, S., Ortega-Mora, L. M., & Saeij, J. P. J. (2018). Toxoplasma CRISPR/Cas9 constructs are functional for gene disruption in Neospora caninum. International journal for parasitology, 48(8), 597–600. https://doi.org/10.1016/j.ijpara.2018.03.0020020-751910.1016/j.ijpara.2018.03.002https://hdl.handle.net/20.500.14352/96373Herein we describe, to our knowledge for the first time the use of the clustered regularly interspaced short palindromic repeats/CRISPR-associated gene 9 (CRISPR/Cas9) system for genome editing of Neospora caninum, an apicomplexan parasite considered one of the main causes of abortion in cattle worldwide. By using plasmids containing the CRISPR/Cas9 components adapted to the closely related parasite Toxoplasma gondii, we successfully knocked out a green fluorescent protein (GFP) in an Nc-1 GFP-expressing strain, and efficiently disrupted the NcGRA7 gene in the Nc-Spain7 isolate by insertion of a pyrimethamine resistance cassette. The successful use of this technology in N. caninum lays the foundation for an efficient, targeted gene modification tool in this parasite.engAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/journal articlehttps://doi.org/10.1016/j.ijpara.2018.03.00229625127restricted accessCRISPR/Cas9Gene disruptionNc-Spain7NcGRA7Neospora caninumTransfectionParasitología (Veterinaria)2415.01 Biología Molecular de Microorganismos