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

被引：17

作者：

Arranz-Solis, David ^{[1
]}

Regidor-Cerrillo, Javier ^{[2
]}

Lourido, Sebastian ^{[3
]}

Miguel Ortega-Mora, Luis ^{[2
]}

Saeij, Jeroen P. J. ^{[1
]}

机构：

[1] Univ Calif Davis, Sch Vet Med, Dept Pathol Microbiol & Immunol, One Shields Ave, Davis, CA 95616 USA

[2] Univ Complutense Madrid, Vet Fac, Anim Hlth Dept, SALUVET, Avda Puerta Hierro S-N, Madrid 28040, Spain

[3] Whitehead Inst Biomed Res, Cambridge, MA 02140 USA

来源：

INTERNATIONAL JOURNAL FOR PARASITOLOGY | 2018年 / 48卷 / 08期

关键词：

Neospora caninum; CRISPR/Cas9; Transfection; Gene disruption; NcGRA7; Nc-Spain7; GONDII; EXPRESSION; MUTATIONS; PROTEINS; TOOL;

D O I：

10.1016/j.ijpara.2018.03.002

中图分类号：

R38 [医学寄生虫学]; Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ; 100103 ;

摘要：

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. (C) 2018 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.

引用

页码：597 / 600

页数：4

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