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Cell Host Microbe. 2019 Oct 9;26(4):478-492.e8. doi: 10.1016/j.chom.2019.09.008.

In Vivo CRISPR Screen Identifies TgWIP as a Toxoplasma Modulator of Dendritic Cell Migration.

Author information

1
Department of Pathology, Microbiology, and Immunology, University of California, Davis, Davis, CA 95615, USA.
2
Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, S-106 91 Stockholm, Sweden.
3
Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA.
4
Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
5
Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA; Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
6
Department of Pathology, Microbiology, and Immunology, University of California, Davis, Davis, CA 95615, USA. Electronic address: jsaeij@ucdavis.edu.

Abstract

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 TgWIP, whose deletion affects Toxoplasma dissemination to distant organs. We show that TgWIP 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. TgWIP affects the morphology of dendritic cells and mediates the dissolution of podosomes, which dendritic cells use to adhere to extracellular matrix. TgWIP 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.

KEYWORDS:

CRISPR; Toxoplasma gondii; WAVE complex; actin; cyst; dendritic cell motility; dissemination; loss-of-function screen; migration; virulence

PMID:
31600500
DOI:
10.1016/j.chom.2019.09.008

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