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dc.contributor.authorRathnapala, UL
dc.contributor.authorGoodman, CD
dc.contributor.authorMcFadden, GI
dc.date.accessioned2021-02-04T01:56:45Z
dc.date.available2021-02-04T01:56:45Z
dc.date.issued2017-06-01
dc.identifierpii: PPATHOGENS-D-17-00352
dc.identifier.citationRathnapala, U. L., Goodman, C. D. & McFadden, G. I. (2017). A novel genetic technique in Plasmodium berghei allows liver stage analysis of genes required for mosquito stage development and demonstrates that de novo heme synthesis is essential for liver stage development in the malaria parasite. PLOS PATHOGENS, 13 (6), https://doi.org/10.1371/journal.ppat.1006396.
dc.identifier.issn1553-7366
dc.identifier.urihttp://hdl.handle.net/11343/259526
dc.description.abstractThe combination of drug resistance, lack of an effective vaccine, and ongoing conflict and poverty means that malaria remains a major global health crisis. Understanding metabolic pathways at all parasite life stages is important in prioritising and targeting novel anti-parasitic compounds. The unusual heme synthesis pathway of the rodent malaria parasite, Plasmodium berghei, requires eight enzymes distributed across the mitochondrion, apicoplast and cytoplasm. Deletion of the ferrochelatase (FC) gene, the final enzyme in the pathway, confirms that heme synthesis is not essential in the red blood cell stages of the life cycle but is required to complete oocyst development in mosquitoes. The lethality of FC deletions in the mosquito stage makes it difficult to study the impact of these mutations in the subsequent liver stage. To overcome this, we combined locus-specific fluorophore expression with a genetic complementation approach to generate viable, heterozygous oocysts able to produce a mix of FC expressing and FC deficient sporozoites. These sporozoites show normal motility and can invade liver cells, where FC deficient parasites can be distinguished by fluorescence and phenotyped. Parasites lacking FC exhibit a severe growth defect within liver cells, with development failure detectable in the early to mid stages of liver development in vitro. FC deficient parasites could not complete liver stage development in vitro nor infect naïve mice, confirming liver stage arrest. These results validate the heme pathway as a potential target for prophylactic drugs targeting liver stage parasites. In addition, we demonstrate that our simple genetic approach can extend the phenotyping window beyond the insect stages, opening considerable scope for straightforward reverse genetic analysis of genes that are dispensable in blood stages but essential for completing mosquito development.
dc.languageEnglish
dc.publisherPUBLIC LIBRARY SCIENCE
dc.rights.urihttps://creativecommons.org/licenses/by/4.0
dc.titleA novel genetic technique in Plasmodium berghei allows liver stage analysis of genes required for mosquito stage development and demonstrates that de novo heme synthesis is essential for liver stage development in the malaria parasite
dc.typeJournal Article
dc.identifier.doi10.1371/journal.ppat.1006396
melbourne.affiliation.departmentSchool of BioSciences
melbourne.affiliation.facultyScience
melbourne.source.titlePLoS Pathogens
melbourne.source.volume13
melbourne.source.issue6
melbourne.identifier.nhmrc1106213
melbourne.identifier.arcDP160104980
dc.rights.licenseCC BY
melbourne.elementsid1215141
melbourne.contributor.authorMcFadden, Geoffrey
melbourne.contributor.authorGoodman, Christopher
melbourne.contributor.authorGallallalage, Upeksha
dc.identifier.eissn1553-7374
melbourne.identifier.fundernameidNHMRC, 1106213
melbourne.identifier.fundernameidAustralian Research Council, DP160104980
melbourne.accessrightsOpen Access


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