An enhanced toolkit for the generation of knockout and marker-free fluorescent Plasmodium chabaudi Article Swipe
YOU?
·
· 2020
· Open Access
·
· DOI: https://doi.org/10.12688/wellcomeopenres.15587.2
· OA: W3016641517
<ns4:p>The rodent parasite <ns4:italic>Plasmodium chabaudi </ns4:italic>is an important <ns4:italic>in vivo </ns4:italic>model of malaria. The ability to produce chronic infections makes it particularly useful for investigating the development of anti-<ns4:italic>Plasmodium</ns4:italic> immunity, as well as features associated with parasite virulence during both the acute and chronic phases of infection. <ns4:italic>P. chabaudi</ns4:italic> also undergoes asexual maturation (schizogony) and erythrocyte invasion in culture, so offers an experimentally-amenable <ns4:italic>in vivo </ns4:italic>to<ns4:italic> in vitro </ns4:italic>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 <ns4:italic>P. chabaudi</ns4:italic> 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 <ns4:italic>Plasmo</ns4:italic>GEM vectors that have revolutionised the scale of experimental genetics in another rodent malaria parasite and used these to generate barcoded <ns4:italic>P. chabaudi </ns4:italic>gene-deletion and –tagging vectors for transfection in our fluorescent <ns4:italic>P. chabaudi</ns4:italic> mother-lines. This produces a tool-kit of <ns4:italic>P. chabaudi </ns4:italic>lines, vectors and transfection approaches that will be of broad utility to the research community.</ns4:p>
