The Mooney Lab

The University of Edinburgh

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


Journal article


E. J. Marr, R. Milne, B. Anar, Gareth Girling, F. Schwach, Jason P Mooney, W. Nahrendorf, P. Spence, Deirdre A. Cunningham, David Baker, J. Langhorne, J. Rayner, O. Billker, Ellen S C Bushell, Joanne Thompson
Wellcome open research, 2020

Semantic Scholar DOI PubMed
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APA   Click to copy
Marr, E. J., Milne, R., Anar, B., Girling, G., Schwach, F., Mooney, J. P., … Thompson, J. (2020). An enhanced toolkit for the generation of knockout and marker-free fluorescent Plasmodium chabaudi. Wellcome Open Research.


Chicago/Turabian   Click to copy
Marr, E. J., R. Milne, B. Anar, Gareth Girling, F. Schwach, Jason P Mooney, W. Nahrendorf, et al. “An Enhanced Toolkit for the Generation of Knockout and Marker-Free Fluorescent Plasmodium Chabaudi.” Wellcome open research (2020).


MLA   Click to copy
Marr, E. J., et al. “An Enhanced Toolkit for the Generation of Knockout and Marker-Free Fluorescent Plasmodium Chabaudi.” Wellcome Open Research, 2020.


BibTeX   Click to copy

@article{e2020a,
  title = {An enhanced toolkit for the generation of knockout and marker-free fluorescent Plasmodium chabaudi.},
  year = {2020},
  journal = {Wellcome open research},
  author = {Marr, E. J. and Milne, R. and Anar, B. and Girling, Gareth and Schwach, F. and Mooney, Jason P and Nahrendorf, W. and Spence, P. and Cunningham, Deirdre A. and Baker, David and Langhorne, J. and Rayner, J. and Billker, O. and Bushell, Ellen S C and Thompson, Joanne}
}

Abstract

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.


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