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Series GSE145800 Query DataSets for GSE145800
Status Public on Feb 25, 2020
Title Differential impacts on host transcription by ROP and GRA effectors from the intracellular parasite Toxoplasma gondii
Organism Toxoplasma gondii RH
Experiment type Expression profiling by high throughput sequencing
Summary The intracellular parasite Toxoplasma gondii employs a vast array of effector proteins from the rhoptry and dense granule organelles to modulate host cell biology; these effectors are known as ROPs and GRAs, respectively. To examine the individual impacts of ROPs and GRAs on host gene expression, we developed a robust, novel protocol to enrich for ultra-pure populations of a naturally occurring and reproducible population of host cells called uninfected-injected (U-I) cells, which Toxoplasma injects with ROPs but subsequently fails to invade. We then performed single cell transcriptomic analysis at 1-3 hours post-infection on U-I cells (as well as on uninfected and infected controls) arising from infection with either wild type parasites or parasites lacking the MYR1 protein, which is required for soluble GRAs to cross the parasitophorous vacuole membrane (PVM) and reach the host cell cytosol. Based on comparisons of infected and U-I cells, the host’s earliest response to infection appears to be driven primarily by the injected ROPs, which appear to induce immune and cellular stress pathways. These ROP-dependent pro-inflammatory signatures appear to be counteracted by at least some of the MYR1-dependent GRAs and may be enhanced by the MYR-independent GRAs, (which are found embedded within the PVM). Finally, signatures detected in uninfected bystander cells from the infected monolayers suggests that MYR1-dependent paracrine effects also counteract inflammatory ROP-dependent processes.
 
Overall design A total of 8395 cells were sequenced and a total of 5354 cells passed quality control and were sequenced. Samples were treated for three timepoints. For each treatment, we have measured a negative control sample treated with mock infection.
 
Contributor(s) Rastogi S, Xue Y, Quake SR, Boothroyd JC
Citation(s) 32518180
Submission date Feb 24, 2020
Last update date Jul 27, 2020
Contact name Yuan Xue
E-mail(s) xuesoso@gmail.com
Organization name Stanford University
Street address 318 Campus Drive
City Stanford
State/province CA
ZIP/Postal code 94305
Country USA
 
Platforms (1)
GPL28187 Illumina NovaSeq 6000 (Toxoplasma gondii RH)
Samples (23)
GSM4333096 1009902407
GSM4333097 1009902401
GSM4333098 1009902404
Relations
BioProject PRJNA608271
SRA SRP250474

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Supplementary file Size Download File type/resource
GSE145800_GEO_deposit_supplementary.tar.gz 471.3 Mb (ftp)(http) TAR
GSE145800_ad_mm_full.h5ad.hdf5 262.4 Mb (ftp)(http) HDF5
GSE145800_adata_raw_mm.h5ad.hdf5 139.6 Mb (ftp)(http) HDF5
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Raw data are available in SRA
Processed data are available on Series record

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