Project description:mRNA profiles of cysts from Entamoeba invadens treatment with ethanol and treament with 200 nM of Trichostatin A during 24h, using Illumina NextSeq 500
Project description:RNA-seq was performed on encysting and excysting E. invadens, a parasite of reptiles that is used as a model system to study stage conversion in Entamoeba species, including the human pathogen E. histolytica. The goal of the project was to identify changes to the transcriptome during development, in order to better understand the mechanisms of development.
Project description:The developmental life cycle of the enteric parasite Entamoeba invadens: transcriptome analysis reveals a crucial role for phospholipase D in stage conversion
Project description:We analyzed ~27nt small RNAs from Entamoeba invadens trophozoites, 24h cysts, 72h cysts, and excysting cells (8h) E. invadens trophozoites were induced to encyst by incubation in low glucose media, and parasites harvested at 0, 24 and 72h. A subset of the 72h cysts were induced to excyst, and parasites harvested at 8h. Total RNA was extracted for each sampleand small RNA libraries constructed and sequenced as below
Project description:RNA-seq was performed on encysting and excysting E. invadens, a parasite of reptiles that is used as a model system to study stage conversion in Entamoeba species, including the human pathogen E. histolytica. The goal of the project was to identify changes to the transcriptome during development, in order to better understand the mechanisms of development. RNA was isolated from trophozoites, from encysting parasites (at 8, 24, 48 and 72 hours after transfer to encystation media) and from excysting parasites (2 and 8 hours after induction of excystation), converted to cDNA, and sequenced on a SOLiD platform. RNA-seq data was used to refine gene models, find potential unannotated genes, and identify genes that are developmentally regulated.
Project description:Entamoeba histolytica is a protozoan parasite that causes colitis and liver abscesses. Several Entamoeba species and strains with differing levels of virulence have been identified. E. histolytica HM-1:IMSS is a virulent strain, E. histolytica Rahman is a nonvirulent strain, and Entamoeba dispar is a nonvirulent species. We used an E. histolytica DNA microarray consisting of 2,110 genes to assess the transcriptional differences between these species/strains with the goal of identifying genes whose expression correlated with a virulence phenotype. We found 415 genes expressed at lower levels in E. dispar and 32 genes with lower expression in E. histolytica Rahman than in E. histolytica HM-1:IMSS. Overall, 29 genes had decreased expression in both the nonvirulent species/strains than the virulent E. histolytica HM-1:IMSS. Interestingly, a number of genes with potential roles in stress response and virulence had decreased expression in either one or both nonvirulent Entamoeba species/strains. These included genes encoding Fe hydrogenase (9.m00419), peroxiredoxin (176.m00112), type A flavoprotein (6.m00467), lysozyme (6.m00454), sphingomyelinase C (29.m00231), and a hypothetical protein with homology to both a Plasmodium sporozoite threonine-asparagine-rich protein (STARP) and a streptococcal hemagglutinin (238.m00054). The function of these genes in Entamoeba and their specific roles in parasite virulence need to be determined. We also found that a number of the non-long-terminal-repeat retrotransposons (EhLINEs and EhSINEs), which have been shown to modulate gene expression and genomic evolution, had lower expression in the nonvirulent species/strains than in E. histolytica HM-1:IMSS. Our results, identifying expression profiles and patterns indicative of a virulence phenotype, may be useful in characterizing the transcriptional framework of virulence. A species experiment design type assays differences between distinct species. Keywords: species_design
