A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes

被引：554

作者：

Sidik, Saima M. ^{[1
]}

Huet, Diego ^{[1
]}

Ganesan, Suresh M. ^{[2
]}

Huynh, My-Hang ^{[3
]}

Wang, Tim ^{[1
,4
,5
]}

Nasamu, Armiyaw S. ^{[2
]}

Thiru, Prathapan ^{[1
]}

Saeij, Jeroen P. J. ^{[6
]}

Carruthers, Vern B. ^{[3
]}

Niles, Jacquin C. ^{[2
]}

Lourido, Sebastian ^{[1
]}

机构：

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

[2] MIT, Dept Biol Engn, Cambridge, MA 02139 USA

[3] Univ Michigan, Sch Med, Dept Microbiol & Immunol, Ann Arbor, MI 48109 USA

[4] MIT, Dept Biol, Cambridge, MA 02139 USA

[5] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[6] Univ Calif Davis, Dept Pathol Microbiol & Immunol, Davis, CA 95616 USA

来源：

CELL | 2016年 / 166卷 / 06期

关键词：

MITOCHONDRIAL METABOLISM; MICRONEME DISCHARGE; GONDII; EXPRESSION; PARASITE; MUTATIONS; VIRULENCE; TRANSFECTION; PREDICTION; PROTEINS;

D O I：

10.1016/j.cell.2016.08.019

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Apicomplexan parasites are leading causes of human and livestock diseases such as malaria and toxoplasmosis, yet most of their genes remain uncharacterized. Here, we present the first genome-wide genetic screen of an apicomplexan. We adapted CRISPR/Cas9 to assess the contribution of each gene from the parasite Toxoplasma gondii during infection of human fibroblasts. Our analysis defines similar to 200 previously uncharacterized, fitness-conferring genes unique to the phylum, from which 16 were investigated, revealing essential functions during infection of human cells. Secondary screens identify as an invasion factor the claudin-like apicomplexan microneme protein (CLAMP), which resembles mammalian tight-junction proteins and localizes to secretory organelles, making it critical to the initiation of infection. CLAMP is present throughout sequenced apicomplexan genomes and is essential during the asexual stages of the malaria parasite Plasmodium falciparum. These results provide broad-based functional information on T. gondii genes and will facilitate future approaches to expand the horizon of antiparasitic interventions.

引用

页码：1423 / +

页数：25

共 63 条

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