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

Saima M. Sidik, Diego Huet, Suresh M. Ganesan, My Hang Huynh, Tim Wang, Armiyaw S. Nasamu, Prathapan Thiru, Jeroen Saeij, Vern B. Carruthers, Jacquin C. Niles, Sebastian Lourido

Research output: Contribution to journalArticle

147 Citations (Scopus)

Abstract

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 ∼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.

Original languageEnglish (US)
Pages (from-to)1423-1435.e12
JournalCell
Volume166
Issue number6
DOIs
StatePublished - Sep 8 2016

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Essential Genes
Toxoplasma
Genes
Genome
Parasites
Toxoplasmosis
Tight Junction Proteins
Antiparasitic Agents
Falciparum Malaria
Livestock
Infection
Organelles
Malaria
Proteins
Fibroblasts
Agriculture
Cells

Keywords

  • Apicomplexan parasites
  • eukaryotic pathogen
  • genome-wide CRISPR screen
  • host-cell invasion
  • host-pathogen interactions
  • malaria
  • toxoplasmosis

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Sidik, S. M., Huet, D., Ganesan, S. M., Huynh, M. H., Wang, T., Nasamu, A. S., ... Lourido, S. (2016). A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Cell, 166(6), 1423-1435.e12. https://doi.org/10.1016/j.cell.2016.08.019

A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. / Sidik, Saima M.; Huet, Diego; Ganesan, Suresh M.; Huynh, My Hang; Wang, Tim; Nasamu, Armiyaw S.; Thiru, Prathapan; Saeij, Jeroen; Carruthers, Vern B.; Niles, Jacquin C.; Lourido, Sebastian.

In: Cell, Vol. 166, No. 6, 08.09.2016, p. 1423-1435.e12.

Research output: Contribution to journalArticle

Sidik, SM, Huet, D, Ganesan, SM, Huynh, MH, Wang, T, Nasamu, AS, Thiru, P, Saeij, J, Carruthers, VB, Niles, JC & Lourido, S 2016, 'A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes', Cell, vol. 166, no. 6, pp. 1423-1435.e12. https://doi.org/10.1016/j.cell.2016.08.019
Sidik SM, Huet D, Ganesan SM, Huynh MH, Wang T, Nasamu AS et al. A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Cell. 2016 Sep 8;166(6):1423-1435.e12. https://doi.org/10.1016/j.cell.2016.08.019
Sidik, Saima M. ; Huet, Diego ; Ganesan, Suresh M. ; Huynh, My Hang ; Wang, Tim ; Nasamu, Armiyaw S. ; Thiru, Prathapan ; Saeij, Jeroen ; Carruthers, Vern B. ; Niles, Jacquin C. ; Lourido, Sebastian. / A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. In: Cell. 2016 ; Vol. 166, No. 6. pp. 1423-1435.e12.
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