High-throughput identification of Toxoplasma gondii effector proteins that target host cell transcription

Simon Butterworth, Kristina Kordova, Sambamurthy Chandrasekaran, Kaitlin K. Thomas, Francesca Torelli, Eloise J. Lockyer, Amelia Edwards, Robert Goldstone, Anita A. Koshy, Moritz Treeck

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Intracellular pathogens and other endosymbionts reprogram host cell transcription to suppress immune responses and recalibrate biosynthetic pathways. This reprogramming is critical in determining the outcome of infection or colonization. We combine pooled CRISPR knockout screening with dual host-microbe single-cell RNA sequencing, a method we term dual perturb-seq, to identify the molecular mediators of these transcriptional interactions. Applying dual perturb-seq to the intracellular pathogen Toxoplasma gondii, we are able to identify previously uncharacterized effector proteins and directly infer their function from the transcriptomic data. We show that TgGRA59 contributes to the export of other effector proteins from the parasite into the host cell and identify an effector, TgSOS1, that is necessary for sustained host STAT6 signaling and thereby contributes to parasite immune evasion and persistence. Together, this work demonstrates a tool that can be broadly adapted to interrogate host-microbe transcriptional interactions and reveal mechanisms of infection and immune evasion.

Original languageEnglish (US)
Pages (from-to)1748-1762.e8
JournalCell Host and Microbe
Volume31
Issue number10
DOIs
StatePublished - Oct 11 2023

Keywords

  • CRISPR screening
  • STAT6
  • Toxoplasma gondii
  • effector proteins
  • host-microbe interactions
  • host-pathogen interactions
  • immune evasion
  • perturb-seq
  • single-cell RNA sequencing

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Virology

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