CRISPR-Cas9-based genome-wide screening of Toxoplasma gondii.

ABSTRACT: Apicomplexan parasites, such as Toxoplasma gondii, cause extensive morbidity and mortality in humans and livestock, highlighting the need for a deeper understanding of their molecular biology. Although techniques for the generation of targeted gene disruptions have long been available for apicomplexans, such methods are not readily scalable to the entire genome. We recently used CRISPR-Cas9 to disrupt all nuclear protein-coding genes in T. gondii using a pooled format. The method relies on transfection of a guide RNA library into parasites constitutively expressing Cas9. Here, we present the complete workflow of such a screen, including preparation of the guide RNA library, growth and testing of the recipient strain, generation of the mutant population, culture conditions for the screen, p

SUBMITTER: Sidik SM 

PROVIDER: S-EPMC6548566 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature