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


ABSTRACT: 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.

SUBMITTER: Arranz-Solis D 

PROVIDER: S-EPMC7025762 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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

Arranz-Solís David D   Regidor-Cerrillo Javier J   Lourido Sebastian S   Ortega-Mora Luis Miguel LM   Saeij Jeroen P J JPJ  

International journal for parasitology 20180403 8


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-e  ...[more]

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