A tetracycline-repressible transactivator system to study essential genes in malaria parasites

Paco Pino, Sarah Sebastian, Eunbin Arin Kim, Erin Bush, Mathieu Brochet, Katrin Volkmann, Elyse Kozlowski, Manuel Llinás, Oliver Billker, Dominique Soldati-Favre

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

A major obstacle in analyzing gene function in apicomplexan parasites is the absence of a practical regulatable expression system. Here, we identified functional transcriptional activation domains within Apicomplexan AP2 (ApiAP2) family transcription factors. These ApiAP2 transactivation domains were validated in blood-, liver-, and mosquito-stage parasites and used to create a robust conditional expression system for stage-specific, tetracycline-dependent gene regulation in Toxoplasma gondii, Plasmodium berghei, and Plasmodium falciparum. To demonstrate the utility of this system, we created conditional knockdowns of two essential P. berghei genes: profilin (PRF), a protein implicated in parasite invasion, and N-myristoyltransferase (NMT), which catalyzes protein acylation. Tetracycline-induced repression of PRF and NMT expression resulted in a dramatic reduction in parasite viability. This efficient regulatable system will allow for the functional characterization of essential proteins that are found in these important parasites.

Original languageEnglish (US)
Pages (from-to)824-834
Number of pages11
JournalCell Host and Microbe
Volume12
Issue number6
DOIs
StatePublished - Dec 13 2012

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Microbiology
  • Virology

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    Pino, P., Sebastian, S., Kim, E. A., Bush, E., Brochet, M., Volkmann, K., Kozlowski, E., Llinás, M., Billker, O., & Soldati-Favre, D. (2012). A tetracycline-repressible transactivator system to study essential genes in malaria parasites. Cell Host and Microbe, 12(6), 824-834. https://doi.org/10.1016/j.chom.2012.10.016