%0 Journal Article
%A Arranz Solís, David
%A Regidor-Cerrillo, Javier
%A Lourido, Sebastian
%A Ortega Mora, Luis Miguel
%A Saeij, Jeroen
%T Toxoplasma CRISPR/Cas9 constructs are functional for gene disruption in Neospora caninum
%D 2018
%@ 0020-7519
%U https://hdl.handle.net/20.500.14352/96373
%X Herein 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.
%~