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Plasmodium berghei Gametocytogenesis

ABSTRACT: Transcriptional profiling of gametocyte non-producer lines in Plasmodium berghei Transcriptome of gametocyte non producer lines (natural and genetic KO) and parental (820) lines. The aim of the study was to identify key genes involved in the decision to commit to gametocytogenesis in Plasmodium berghei. These microarrays compare naturally selected lines that do not produce gametocytes, and the parental line and additionally a genetic knock out of AP2-G PBANKA_143750. Data published Sinha, Hughes, et, al Nature tbc.

ORGANISM(S): Plasmodium berghei

PROVIDER: GSE53246 | GEO | 2013/12/13

SECONDARY ACCESSION(S): PRJNA231246

REPOSITORIES: GEO

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Plasmodium berghei Gametocytogenesis

Project description:Transcriptional profiling of gametocyte non-producer lines in Plasmodium berghei Transcriptome of gametocyte non producer lines (natural and genetic KO) and parental (820) lines. The aim of the study was to identify key genes involved in the decision to commit to gametocytogenesis in Plasmodium berghei. These microarrays compare naturally selected lines that do not produce gametocytes, and the parental line and additionally a genetic knock out of AP2-G PBANKA_143750. Data published Sinha, Hughes, et, al Nature tbc. 2- colour microarray comparing to common background pool (containing all life cycle stages). Replicates of different life cycle stages of gametocyte non-producer lines and wild tye (WT) parental control lines

2013-12-13 | E-GEOD-53246 | biostudies-arrayexpress

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Project description:During the malaria infection, Plasmodium parasites invade the host’s red blood cells where they can differentiate into two different life forms. The majority will replicate asexually and infect new erythrocytes. A small percentage, however, will transform into gametocytes – a specialized sexual stage able to survive and develop when taken up by Anopheles mosquito. As the gametocytes ensure the parasite’s transmission to a new host, their generation is an attractive target for new antimalarial interventions. The molecular mechanisms controlling gametocytogenesis, however, remain largely unknown due to the technical challenges: the early gametocytes are morphologically indistinguishable from asexual parasites and present in very low numbers during the infection. Recently, AP2-G - a transcription factor from an apicomplexa-specific apiAP2 family – was described as indispensable for gametocyte commitment in both human malaria parasite Plasmodium falciparum and rodent malaria model Plasmodium berghei. Therefore, we have decided to test whether the overexpression of this factor alone could increase gametocyte production and enable the investigation of uncharacterised, earliest stages of gametocyte development. To this end, we have engineered PBGAMi - a Plasmodium berghei line, in which all parasites were ap2-g deficient by default but able to overexpress it when induced with rapamycin. While the control parasites (PBGAMi R-), as expected, differentiated into asexual forms (schizonts) only, almost all rapamycin-treated parasites (PBGAMi R+) transformed into gametocytes. We used the generated line to perform RNA-seq analysis of the R- and R+ populations at different time points of their development and identify the changes arising between them, mapping the sequence of events leading to the formation of gametocytes.

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RNA sequencing of two protein phosphatases(ppm2 and ppm5) knockouts in Plasmodium berghei to study their role in the parasite's sexual development

Project description:To identify protein phosphatases (PP) that are essential during asexual blood stage development in Plasmodium berghei, we attempted systematic deletion of the 30 P. berghei PP genes using double homologous recombination. Of these, we performed strand-specific RNA-sequencing (RNA-Seq) analysis for 2 myristoylated PP mutants _ppm2 and _ppm5 lines across relevant lifestages ( activated gametocyte and schizont stages in both _ppm2 and _ppm5 and ookinete stage in _ppm5 alone). Two to four biological replicates each for _ppm2, _ppm5 and wild-type parasites were processed into strand-specific RNA-seq libraries and sequenced in Illumina Hiseq platform using 100-bp paired end chemistry.

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