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PLoS One. 2014 Nov 13;9(11):e112571. doi: 10.1371/journal.pone.0112571. eCollection 2014.

Plasmodium falciparum transfected with ultra bright NanoLuc luciferase offers high sensitivity detection for the screening of growth and cellular trafficking inhibitors.

Author information

1
Macfarlane Burnet Institute of Medical Research and Public Health, Melbourne, Victoria, Australia.
2
Macfarlane Burnet Institute of Medical Research and Public Health, Melbourne, Victoria, Australia; Monash University, Melbourne, Australia.
3
Macfarlane Burnet Institute of Medical Research and Public Health, Melbourne, Victoria, Australia; Monash University, Melbourne, Australia; University of Melbourne, Melbourne, Australia.

Abstract

Drug discovery is a key part of malaria control and eradication strategies, and could benefit from sensitive and affordable assays to quantify parasite growth and to help identify the targets of potential anti-malarial compounds. Bioluminescence, achieved through expression of exogenous luciferases, is a powerful tool that has been applied in studies of several aspects of parasite biology and high throughput growth assays. We have expressed the new reporter NanoLuc (Nluc) luciferase in Plasmodium falciparum and showed it is at least 100 times brighter than the commonly used firefly luciferase. Nluc brightness was explored as a means to achieve a growth assay with higher sensitivity and lower cost. In addition we attempted to develop other screening assays that may help interrogate libraries of inhibitory compounds for their mechanism of action. To this end parasites were engineered to express Nluc in the cytoplasm, the parasitophorous vacuole that surrounds the intraerythrocytic parasite or exported to the red blood cell cytosol. As proof-of-concept, these parasites were used to develop functional screening assays for quantifying the effects of Brefeldin A, an inhibitor of protein secretion, and Furosemide, an inhibitor of new permeation pathways used by parasites to acquire plasma nutrients.

PMID:
25392998
PMCID:
PMC4231029
DOI:
10.1371/journal.pone.0112571
[Indexed for MEDLINE]
Free PMC Article

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