| Full text | |
| Author(s): |
Lima, Caroline
;
Verdaguer, Ignasi B.
;
Wunderlich, Gerhard
;
Katzin, Alejandro M.
;
Crabb, Brendan S.
;
Gilson, Paul R.
;
Azevedo, Mauro F.
Total Authors: 7
|
| Document type: | Journal article |
| Source: | Experimental Parasitology; v. 254, p. 8-pg., 2023-11-01. |
| Abstract | |
Conditional gene expression is a powerful tool to investigate putative vaccine and drug targets, especially in a haploid organism such as Plasmodium falciparum. Inducible systems based on regulation of either transcription, translation, protein or mRNA stability, among others, allow switching on an off the expression of any desired gene causing specific gain or loss of function phenotypes. However, those systems can be cumbersome involving the construction of large plasmids and generation of multiple transgenic parasite lines. In addition, the dynamic range of regulation achieved is not predictable for each individual gene and can be insufficient to generate detectable phenotypes when the genes of interest are silenced. Here, we combined up to three distinct inducible systems to regulate the expression of a single gene. Expression of the reporter NanoLuc luciferase was regulated over 40-fold, which correlates to the regulation achieved by each individual system multiplied by each other. We applied the conditionally expressed NanoLuc to evaluate the effect of fast-acting antimalarials such as chloroquine and artesunate as well as of slower-acting ones such as atovaquone. The conditionally expressed reporter allowed faster and more reliable detection of toxicity to the parasite, which correlated to the expected action of each compound. Bioluminescence achieved by the expression of this inducible highly sensitive reporter is therefore a promising tool to investigate the temporal effect of potential new antimalarials. This single plasmid combination system might also prove useful to achieve sufficient regulation of genes of interest to produce lossof-function phenotypes. (AU) | |
| FAPESP's process: | 15/19316-3 - Regulation of P. falciparum egress from the host cell: identification of new targets |
| Grantee: | Mauro Ferreira de Azevedo |
| Support Opportunities: | Research Grants - Young Investigators Grants |
| FAPESP's process: | 18/08989-5 - Evaluation of the combination of conditional gene expression in P. falciparum for the study of functional genome and potential antimalarials screening |
| Grantee: | Caroline Lima dos Santos |
| Support Opportunities: | Scholarships in Brazil - Master |
| FAPESP's process: | 18/12589-2 - Multi-user equipment approved in grant 15/19316-3: flow cytometer |
| Grantee: | Mauro Ferreira de Azevedo |
| Support Opportunities: | Multi-user Equipment Program |