Efficient Genome Engineering of Toxoplasma gondii Using CRISPR/Cas9

被引：176

作者：

Sidik, Saima M. ^{[1
]}

Hackett, Caroline G. ^{[1
]}

Tran, Fanny ^{[2
,3
]}

Westwood, Nicholas J. ^{[2
,3
]}

Lourido, Sebastian ^{[1
]}

机构：

[1] Whitehead Inst Biomed Res, Cambridge, MA 02142 USA

[2] Univ St Andrews, Sch Chem & Biomed Sci Res Complex, St Andrews, Fife, Scotland

[3] EaStCHEM, St Andrews, Fife, Scotland

来源：

PLOS ONE | 2014年 / 9卷 / 06期

关键词：

INVASION; SYSTEMS; MARKER; GENES;

D O I：

10.1371/journal.pone.0100450

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Toxoplasma gondii is a parasite of humans and animals, and a model for other apicomplexans including Plasmodium spp., the causative agents of malaria. Despite many advances, manipulating the T. gondii genome remains labor intensive, and is often restricted to lab-adapted strains or lines carrying mutations that enable selection. Here, we use the RNA-guided Cas9 nuclease to efficiently generate knockouts without selection, and to introduce point mutations and epitope tags into the T. gondii genome. These methods will streamline the functional analysis of parasite genes and enable high-throughput engineering of their genomes.

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页数：8

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