Toxoplasma CRISPR/Cas9 constructs are functional for gene disruption in Neospora caninum

David Arranz-Solís, Javier Regidor-Cerrillo, Sebastian Lourido, Luis Miguel Ortega-Mora, Jeroen Saeij

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)597-600
Number of pages4
JournalInternational Journal for Parasitology
Issue number8
StatePublished - Jul 1 2018


  • CRISPR/Cas9
  • Gene disruption
  • Nc-Spain7
  • NcGRA7
  • Neospora caninum
  • Transfection

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases


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