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A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites
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 wer...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cell Press
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712325/ https://www.ncbi.nlm.nih.gov/pubmed/23245327 http://dx.doi.org/10.1016/j.chom.2012.10.016 |
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author | Pino, Paco Sebastian, Sarah Kim, EunBin Arin Bush, Erin Brochet, Mathieu Volkmann, Katrin Kozlowski, Elyse Llinás, Manuel Billker, Oliver Soldati-Favre, Dominique |
author_facet | Pino, Paco Sebastian, Sarah Kim, EunBin Arin Bush, Erin Brochet, Mathieu Volkmann, Katrin Kozlowski, Elyse Llinás, Manuel Billker, Oliver Soldati-Favre, Dominique |
author_sort | Pino, Paco |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3712325 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Cell Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-37123252013-07-17 A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites Pino, Paco Sebastian, Sarah Kim, EunBin Arin Bush, Erin Brochet, Mathieu Volkmann, Katrin Kozlowski, Elyse Llinás, Manuel Billker, Oliver Soldati-Favre, Dominique Cell Host Microbe Resource 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. Cell Press 2012-12-13 /pmc/articles/PMC3712325/ /pubmed/23245327 http://dx.doi.org/10.1016/j.chom.2012.10.016 Text en © 2012 Elsevier Inc. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license |
spellingShingle | Resource Pino, Paco Sebastian, Sarah Kim, EunBin Arin Bush, Erin Brochet, Mathieu Volkmann, Katrin Kozlowski, Elyse Llinás, Manuel Billker, Oliver Soldati-Favre, Dominique A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites |
title | A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites |
title_full | A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites |
title_fullStr | A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites |
title_full_unstemmed | A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites |
title_short | A Tetracycline-Repressible Transactivator System to Study Essential Genes in Malaria Parasites |
title_sort | tetracycline-repressible transactivator system to study essential genes in malaria parasites |
topic | Resource |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3712325/ https://www.ncbi.nlm.nih.gov/pubmed/23245327 http://dx.doi.org/10.1016/j.chom.2012.10.016 |
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