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Widespread variation in transcript abundance within and across developmental stages of Trypanosoma brucei

ABSTRACT: Whole genome comparison of RNA levels for both protein coding genes and structural RNAs in five different life cycle stages: in vivo slender bloodstream form, in vivo stumpy bloodstream form, cultured bloodstream form, log-phase procyclic culture form and stationary-phase procyclic culture form RNA from three independent biological replicates from five different life cycle stages were hybridized to Nimblegen arrays (Madison,WI USA) that contained 8 probes per open reading frame and 3 probes per structural RNA spotted three times per array

ORGANISM(S): Trypanosoma brucei TREU927

SUBMITTER: Charles Kifer

PROVIDER: E-GEOD-18049 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Widespread variation in transcript abundance within and across developmental stages of Trypanosoma brucei

Project description:Whole genome comparison of RNA levels for both protein coding genes and structural RNAs in five different life cycle stages: in vivo slender bloodstream form, in vivo stumpy bloodstream form, cultured bloodstream form, log-phase procyclic culture form and stationary-phase procyclic culture form

2009-09-24 | GSE18049 | GEO

Dynamic mRNA Expression analysis of cells undergoing synchronous life-cycle differentiation in Trypanosoma brucei

Project description:mRNA expression profiles of trypanosomes from two discrete bloodstream form stages of the parasite (slender and stumpy forms), as well as during the transition of the stumpy population to the procyclic life-cycle stage were studied. Our analysis represents the first comparison of in vivo derived pleomorphic slender cells with genetically identical stumpy forms, and a first analysis of the dynamic changes in mRNA profile that accompany the transition to procyclic forms.

2009-07-10 | GSE17026 | GEO

Dynamic mRNA Expression analysis of cells undergoing synchronous life-cycle differentiation in Trypanosoma brucei

2010-05-16 | E-GEOD-17026 | biostudies-arrayexpress

Phenotypic activation and repression of three ELAV-like proteins in Trypanosoma brucei

Project description:Three potential ELAV-like proteins of T. brucei, including Tb927.3.2930, Tb927.7.5380, and Tb927.8.6650, were either inhibited by RNAi or phenotypically activated by over-expression, followed by microarray analysis of the transcriptome. The results indicated that these ELAV-like proteins regulate the abundance of a large number of T. brucei transcripts, potentially through regulation of mRNA stability. Each of the three ELAV-like proteins were either inhibited by RNAi or over-expressed, in stable transgenic procyclic form cell lines. Total RNA was extracted 48h after tetracycline induction of the constructs (except for total RNA from Tb927.8.6650 RNAi which was extracted 24h after tet-induction), and sent to NimbleGen for cDNA synthesis and hybridization. Non-induced cells were analyzed in parallel.

2013-04-18 | E-GEOD-46161 | biostudies-arrayexpress

Regulation of Trypanosoma brucei total and polysomal mRNA during development within its mammalian host

Project description:The gene expression of Trypanosoma brucei has been examined extensively in the blood of mammalian hosts and in forms found in the midgut of its arthropod vector, the tsetse fly. However, trypanosomes also undergo development within the mammalian bloodstream as they progress from morphologically M-bM-^@M-^Xslender formsM-bM-^@M-^Y to transmissible M-bM-^@M-^Xstumpy formsM-bM-^@M-^Y through morphological intermediates. This transition is temporally progressive within the first wave of parasitaemia such that gene expression can be monitored in relatively pure slender and stumpy populations as well as during the progression between these extremes. The development also represents the progression of cells from translationally active forms adapted for proliferation in the host to translationally quiescent forms, adapted for transmission. We have used metabolic labelling to quantitate translational activity in slender forms, stumpy forms and in forms undergoing early differentiation to procyclic forms in vitro. Thereafter we have examined the cohort of total mRNAs that are enriched in throughout development in the mammalian bloodstream (slender, intermediate and stumpy forms), irrespective of strain, revealing those that exhibit consistent developmental regulation rather than sample specific changes. Transcripts that cosediment with polysomes in stumpy forms and slender forms have also been identified to enrich transcripts that escape translational repression prior to transmission. Combined, the expression and polysomal association of transcripts as trypanosomes undergo development in the mammalian bloodstream haves been defined, providing a resource for trypanosome researchers. This facilitates the identification of those that undergo developmental regulation in the bloodstream and therefore those likely to have a role in the survival and capacity for transmission of stumpy forms. Examination of gene expression during life cycle stages.

2013-12-16 | E-GEOD-46388 | biostudies-arrayexpress

Life stage specific poly(A) site selection regulated by Trypanosoma brucei DRBD18

Project description:In this work, we examine the function of DRBD18 in T. brucei, here using single cell RNA sequencing (scRNAseq) of uninduced and induced DRBD18 knockdowns. In insect procyclic form, we identified one major and one minor population of cells that dramatically increased upon DRBD18 knockdown. Both of these cell populations were significantly enriched for transcripts that are normally upregulated in the quiescent stumpy bloodstream form and metacyclic (MF) life cycle stages, consistent with a role for DRBD18 in life cycle progression. Furthermore, the 3’ end bias of our sequencing technique allowed us to investigate the effect of DRBD18 on poly(A) site selection, and extension of gene annotations permitted a comprehensive analysis of these changes. We found that 3’UTR shortening upon DRBD18 depletion was prevalent, with over 1500 transcripts exhibiting this this phenotype, and RIPseq analyses showed significant overlap between bound transcripts and those with 3’UTR shortening

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Transcriptome analysis of two life cycle stages of Trypanosoma brucei gambiense

Project description:Trypanosoma brucei gambiense is the causative agent of the fatal human disease African sleeping sickness. Here we have compared the transcriptome of two different life cycle stages, the potentially human-infective bloodstream form and the non-human-infective procyclic stage, using digital gene expression (DGE) analysis. Digital gene expression analysis was performed on RNA from 3 biological replicates of bloodstream cultured T.b. gambiense strain STIB 386 and compared to that from 3 biological replicates of procyclic cultured T.b. gambiense strain STIB 386.

2010-07-21 | E-GEOD-18065 | biostudies-arrayexpress

Transcriptome analysis of two life cycle stages of Trypanosoma brucei gambiense

2010-07-22 | GSE18065 | GEO

Regulation of Trypanosoma brucei total and polysomal mRNA during development within its mammalian host

Project description:The gene expression of Trypanosoma brucei has been examined extensively in the blood of mammalian hosts and in forms found in the midgut of its arthropod vector, the tsetse fly. However, trypanosomes also undergo development within the mammalian bloodstream as they progress from morphologically ‘slender forms’ to transmissible ‘stumpy forms’ through morphological intermediates. This transition is temporally progressive within the first wave of parasitaemia such that gene expression can be monitored in relatively pure slender and stumpy populations as well as during the progression between these extremes. The development also represents the progression of cells from translationally active forms adapted for proliferation in the host to translationally quiescent forms, adapted for transmission. We have used metabolic labelling to quantitate translational activity in slender forms, stumpy forms and in forms undergoing early differentiation to procyclic forms in vitro. Thereafter we have examined the cohort of total mRNAs that are enriched in throughout development in the mammalian bloodstream (slender, intermediate and stumpy forms), irrespective of strain, revealing those that exhibit consistent developmental regulation rather than sample specific changes. Transcripts that cosediment with polysomes in stumpy forms and slender forms have also been identified to enrich transcripts that escape translational repression prior to transmission. Combined, the expression and polysomal association of transcripts as trypanosomes undergo development in the mammalian bloodstream haves been defined, providing a resource for trypanosome researchers. This facilitates the identification of those that undergo developmental regulation in the bloodstream and therefore those likely to have a role in the survival and capacity for transmission of stumpy forms.

2013-12-16 | GSE46388 | GEO

Multiple levels of gene regulation mediate differentiation of the intracellular pathogen Leishmania

Project description:Whole genome high density tiliing array of eight timepoints from the promastitgote to amastigote differentiation process. RNA from two independent biological replicates from eight points in the promastigote to amastigote differentiation process were hybridized to Nimblegen arrays (Madison,WI USA) that contained 10 probes per open reading frame and 3 probes per structural RNA spotted three times per array. The microarray probes were designed to Leishmania infantum strain JCPM5 and the sample RNA was obtained from Leishmania donovani strain S1. The two species are thought to be nearly identical at the nucleotide level, however whole-genome sequence was not available from L. donovani.

2010-08-15 | E-GEOD-21936 | biostudies-arrayexpress

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