Trigger-induced RNAi gene silencing to identify pathogenicity factors of Entamoeba histolytica

FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology
Jenny MatthiesenI Bruchhaus

Abstract

Recently, Entamoeba histolytica clones derived from isolate HM-1:IMSS that differ in their pathogenicity were identified. Whereas some clones induce amoebic liver abscesses (ALAs) in animal models of amoebiasis, others provoke only minimal liver lesions. Based on transcriptome studies of pathogenic and nonpathogenic clones, differentially expressed genes associated with reduced or increased liver pathology can be identified. Here, to analyze the influence of these genes on ALA formation in more detail, an RNA interference-trigger mediated silencing approach was used. Using newly identified trigger sequences, the expression of 15 genes was silenced. The respective transfectants were analyzed for their ability to induce liver destruction in the murine model for the disease. Silencing of EHI_180390 (encoding an AIG1 protein) increased liver pathology induced by a nonpathogenic parent clone, whereas silencing of EHI_127670 (encoding a hypothetical protein) decreased the pathogenicity of an initially pathogenic parent clone. Additional phenotypical in vitro analyses of EHI_127670 silencing as well as overexpression transfectants indicated that this molecule has an influence on size, growth, and cysteine peptidase activity of E. hist...Continue Reading

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Citations

Mar 7, 2020·Genetics and Molecular Biology·Gabriel José de CarliTiago Campos Pereira
Apr 6, 2021·Frontiers in Cellular and Infection Microbiology·Constantin KönigIris Bruchhaus

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