Project description:BackgroundEntamoeba histolytica is the causative agent of amebiasis, a disease that is a major source of morbidity and mortality in the developing world. MicroRNAs (miRNAs) are a large group of non-coding RNAs that play important roles in regulating gene expression and protein translation in animals. Genome-wide identification of miRNAs is a critical step to facilitating our understanding of genome organization, genome biology, evolution, and post-transcriptional regulation.Methodology/principal findingsWe sequenced a small RNA library prepared from a culture of trophozoites of Entamoeba histolytica Strain HM1-IMSS using a deep DNA sequencing approach. Deep sequencing yielded 16 million high-quality short sequence reads containing a total of 5 million non-redundant sequence reads. Based on a bioinformatics pipeline, we found that only 0.5% of these non-redundant small RNA reads were a perfect match with the drafted E. histolytica genome. We did not find miRNA homologs in plant or animal miRNAs. We discovered 199 new potential Entamoeba histolytica miRNAs. The expression and sequence of these Ehi-miRNAs were further validated through microarray by µParaflo Microfluidic Biochip Technology. Ten potential miRNAs were additionally confirmed by real time RT-PCR analysis. Prediction of target genes matched 32 known genes and 34 hypothetical genes.Conclusions/significanceThese results show that there is a number of regulatory miRNAs in Entamoeba histolytica. The collection of miRNAs in this parasite could be used as a new platform to study genomic structure, gene regulation and networks, development, and host-parasite interactions.
Project description:This dataset contains RNA-seq data from Entamoeba histolytica strains, including a KERP2-knockdown strain (psAP-KERP2gs) and a control strain (psAP-mock). To investigate the functional role of KERP2 (EHI_065630), the knockdown strain was generated using small interfering RNAs with the psAP-2-Gunma plasmid. RNA-Seq analysis revealed distinct transcriptional profiles between KERP2gs and psAP-mock strains. GO and KEGG pathway enrichment analyses identified upregulation of genes involved in proteolysis regulation, sulfur amino acid metabolism, and amoebiasis in the KERP2-knockdown strain. Notably, cysteine synthases (EHI_024230, EHI_160930), methionine γ-lyase (EHI_057550), cysteine protease (EHI_010850), and pore-forming peptides (EHI_169350, EHI_194540, EHI_15940) were significantly upregulated. These findings suggest a potential role for KERP2 in regulating parasitic activities, possibly through chromatin binding. This dataset provides a valuable resource for studying KERP2-associated gene expression changes in E. histolytica.