In Vivo  CRISPR Screen Identifies  Tg WIP as a  Toxoplasma  Modulator of Dendritic Cell Migration

• Published in: Cell host & microbe Vol. 26; no. 4; p. 478
• Main Authors: Sangaré, Lamba Omar, Ólafsson, Einar B., Wang, Yifan, Yang, Ninghan, Julien, Lindsay, Camejo, Ana, Pesavento, Patricia, Sidik, Saima M., Lourido, Sebastian, Barragan, Antonio, Saeij, Jeroen P. J.
• Format: Journal Article
• Language: English
• Published: 2019
• Subjects: actin; CRISPR; cyst; dendritic cell motility; dissemination; loss-of-function screen; migration; Molecular Cellbiology; molekylär cellbiologi; Toxoplasma gondii; virulence; WAVE complex
• ISSN: 1931-3128; 1934-6069
• DOI: 10.1016/j.chom.2019.09.008

Toxoplasma  can reach distant organs, especially the brain, leading to a lifelong chronic phase. However, genes involved in related  in vivo  processes are currently unknown. Here, we use focused CRISPR libraries to identify  Toxoplasma  genes that affect  in vivo  fitness. We focus on  Tg WIP, whose deletion affects  Toxoplasma dissemination to distant organs. We show that  Tg WIP is secreted into the host cell upon invasion and interacts with the host WAVE regulatory complex and SHP2 phosphatase, both of which regulate actin dynamics.  Tg WIP affects the morphology of dendritic cells and mediates the dissolution of podosomes, which dendritic cells use to adhere to extracellular matrix.  Tg WIP enhances the motility and transmigration of parasitized dendritic cells, likely explaining its effect on  in vivo fitness. Our results provide a framework for systemic identification of  Toxoplasma genes with  in vivo  effects at the site of infection or on dissemination to distant organs, including the brain.