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An enhanced toolkit...
An enhanced toolkit for the generation of knockout and marker-free fluorescent Plasmodium chabaudi
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Marr, Edward J (author)
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Milne, Rachel M (author)
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Anar, Burcu (author)
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Girling, Gareth (author)
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Schwach, Frank (author)
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Mooney, Jason P (author)
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Nahrendorf, Wiebke (author)
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Spence, Philip J (author)
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Cunningham, Deirdre (author)
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Baker, David A (author)
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Langhorne, Jean (author)
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Rayner, Julian C (author)
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- Billker, Oliver (author)
- Umeå universitet,Institutionen för molekylärbiologi (Teknisk-naturvetenskaplig fakultet),Molekylär Infektionsmedicin, Sverige (MIMS),Parasites and Microbes, Wellcome Sanger Institute, Wellcome Genome Campus, Cambridge, UK
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- Bushell, Ellen S. (author)
- Umeå universitet,Institutionen för molekylärbiologi (Medicinska fakulteten),Molekylär Infektionsmedicin, Sverige (MIMS),Parasites and Microbes, Wellcome Sanger Institute, Wellcome Genome Campus, Cambridge, UK
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Thompson, Joanne (author)
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(creator_code:org_t)
- 2020-04-17
- 2020
- English.
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In: Wellcome open research. - : Wellcome open research. - 2398-502X. ; 5
- Related links:
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https://doi.org/10.1...
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https://doi.org/10.1...
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Abstract
Subject headings
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- 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.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Mikrobiologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Microbiology in the medical area (hsv//eng)
Keyword
- PlasmoGem
- Plasmodium chabaudi
- Transfection
- malaria
- mikrobiologi
- Microbiology
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Marr, Edward J
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Milne, Rachel M
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Anar, Burcu
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Girling, Gareth
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Schwach, Frank
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Mooney, Jason P
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show more...
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Nahrendorf, Wieb ...
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Spence, Philip J
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Cunningham, Deir ...
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Baker, David A
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Langhorne, Jean
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Rayner, Julian C
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Billker, Oliver
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Bushell, Ellen S ...
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Thompson, Joanne
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show less...
- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Microbiology in ...
- Articles in the publication
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Wellcome open re ...
- By the university
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Umeå University