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An enhanced toolkit for the generation of knockout and marker-free fluorescent Plasmodium chabaudi
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2020 (English)In: Wellcome open research, ISSN 2398-502X, Vol. 5, article id 71Article in journal (Refereed) Published
Abstract [en]

The rodent parasite Plasmodium chabaudi is an important in vivo model of malaria. The ability to produce chronic infections makes it particularly useful for investigating the development of anti- Plasmodium immunity, as well as features associated with parasite virulence during both the acute and chronic phases of infection. P. chabaudi also undergoes asexual maturation (schizogony) and erythrocyte invasion in culture, so offers an experimentally-amenable in vivo to in vitro model for studying gene function and drug activity during parasite replication. To extend the usefulness of this model, we have further optimised transfection protocols and plasmids for P. chabaudi and generated stable, fluorescent lines that are free from drug-selectable marker genes. These mother-lines show the same infection dynamics as wild-type parasites throughout the lifecycle in mice and mosquitoes; furthermore, their virulence can be increased by serial blood passage and reset by mosquito transmission. We have also adapted the large-insert, linear PlasmoGEM vectors that have revolutionised the scale of experimental genetics in another rodent malaria parasite and used these to generate barcoded P. chabaudi gene-deletion and -tagging vectors for transfection in our fluorescent P. chabaudi mother-lines. This produces a tool-kit of P. chabaudi lines, vectors and transfection approaches that will be of broad utility to the research community.

Place, publisher, year, edition, pages
Wellcome open research , 2020. Vol. 5, article id 71
Keywords [en]
PlasmoGem, Plasmodium chabaudi, Transfection, malaria
National Category
Microbiology in the medical area
Research subject
Microbiology
Identifiers
URN: urn:nbn:se:umu:diva-172671DOI: 10.12688/wellcomeopenres.15587.1PubMedID: 32500098Scopus ID: 2-s2.0-85086157248OAI: oai:DiVA.org:umu-172671DiVA, id: diva2:1445436
Available from: 2020-06-23 Created: 2020-06-23 Last updated: 2023-03-23Bibliographically approved

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Billker, OliverBushell, Ellen S.

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Billker, OliverBushell, Ellen S.
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Department of Molecular Biology (Faculty of Science and Technology)Molecular Infection Medicine Sweden (MIMS)Department of Molecular Biology (Faculty of Medicine)
Microbiology in the medical area

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CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf