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Genome-wide essential gene identification for Shiga toxin-induced cell death using CRISPR/CAS9 screening system

ABSTRACT: Genome-wide essential gene identification for Shiga toxin-induced cell death using CRISPR/CAS9 screening system

PROVIDER: PRJNA480005 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR7478820.fastq.gz	Fastqsanger.gz
	SRR7478821.fastq.gz	Fastqsanger.gz
	SRR7478814.fastq.gz	Fastqsanger.gz
	SRR7478815.fastq.gz	Fastqsanger.gz
	SRR7478816.fastq.gz	Fastqsanger.gz

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Genome-wide essential gene identification for Shiga toxin-induced cell death using CRISPR/CAS9 screening system with Vero cells

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Genome-wide essential gene identification for Shiga toxin-induced cell death using CRISPR/CAS9 screening system with Vero cells

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Project description:Changes in endothelial phenotype induced by E. coli-derived Shiga toxins (Stx) are believed to play a critical role in the pathogenesis of hemolytic uremic syndrome. Stx inactivate host ribosomes, but also alter gene expression at concentrations that minimally affect global protein synthesis. The effect of Stx on the gene expression profile of human microvascular endothelial cells was examined using the Affymetrix HG-U133A platform. Data were processed using 13 different methods and revealed 369 unique differentially expressed genes, 318 of which were up-regulated and 51 of which were down-regulated. These studies implicated activation of the CXCR4/CXCR7/SDF-1 chemokine pathway in Stx-mediated pathogenesis. Primary human dermal microvascular endothelial cells were treated with vehicle or Shiga toxin (10 fM, 24 h, n = 6) and changes in steady-state mRNA levels were determined by hybridization to Affymetrix HG-U133A arrays

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Genome-wide essential gene identification for SubAB toxin-induced cell death using CRISPR/CAS9 screening system

Project description:Isolation of genes whose knockout confer resistance to SubAB-Induced cell death

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Alterations in gene expression in response to different regimes of Shiga toxin 2

Project description:The microarray data provided here belong to a study that describes two different Shiga toxin (Stx) induced models of hemolytic uremic syndrome (HUS) in mice. Although several rodent models of HUS were published, it still remains difficult to reproduce all clinical features of human HUS. Here, two different Stx2 regimes combined with volume resuscitation were tested in C57BL/6J wild type mice. Animals were euthanized because of kidney injury after 3 or 7 days respectively. Kidneys were removed for histological evaluation and RNA extraction. Kidney injury was confirmed histologically and by laboratory parameters in plasma. Data for the respective vehicle treated groups are also provided here.

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Early termination of the Shiga toxin transcript generates a regulatory small RNA

Project description:Enterohaemorrhagic E. coli is significant human pathogen that causes disease ranging from haemorrhagic colitis to haemolytic uremic syndrome. The later can lead to potentially fatal renal failure and is caused by the release of Shiga toxins that are encoded within lambdoid bacteriophages. The toxins are encoded within the late transcript of the phage and are regulated by anti-termination of the PR’ late promoter during lytic induction of the phage. During lysogeny, the late transcript is prematurely terminated at tR’ immediately downstream of PR’, generating a short RNA that is a by-product of anti-termination regulation. We demonstrate that this short transcript binds the small RNA chaperone Hfq, and is processed into a stable 74 nt regulatory small RNA that we have termed, StxS. StxS activates expression of the general stress response sigma factor, RpoS, through direct interactions with an activating seed sequence within the 5’ UTR. StxS represses expression of Shiga toxin 1 and activation of RpoS promotes high cell density growth under nutrient limiting conditions. Many phages utilise anti-termination to regulate the lytic/lysogenic switch and our results demonstrate that short RNAs generated as a by-product of this regulation can acquire regulatory small RNA features and modulate host fitness.

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