An enhanced toolkit for the generation of knockout and marker-free fluorescent Plasmodium chabaudi

Ellen S. Bushell*, Edward J. Marr, Rachel M. Milne, Burcu Anar, Gareth Girling, Frank Schwach, Jason P. Mooney, Wiebke Nahrendorf, Philip J. Spence, Deirdre Cunningham, David A. Baker, Jean Langhorne, Julian C. Rayner, Oliver Billker, Joanne Thompson

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


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.

Original languageEnglish
Article number71
JournalWellcome Open Research
Publication statusPublished - 2020
Externally publishedYes

Bibliographical note

Funding Information:
Grant information: This work was supported by the Wellcome Trust through a Seed Award in Science to JT [202094] and core funding to

Publisher Copyright:
© 2020 Marr EJ et al.


  • Malaria
  • Plasmodium chabaudi
  • PlasmoGem
  • Transfection


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