A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites

作者：Pino Paco; Sebastian Sarah; Kim EunBin Arin; Bush Erin; Brochet Mathieu; Volkmann Katrin; Kozlowski Elyse; Llinas Manuel; Billker Oliver^*; Soldati Favre Dominique

来源：Cell Host & Microbe, 2012, 12(6): 824-834.

DOI：10.1016/j.chom.2012.10.016

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.

• 出版日期2012-12-13