A Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the <named-content content-type="genus-species">Plasmodium falciparum</named-content> Life Cycle
ABSTRACT Plasmodium falciparum has a complex life cycle that involves interaction with multiple tissues inside the human and mosquito hosts. Identification of essential genes at all different stages of the P. falciparum life cycle is urgently required for clinical development of tools for malaria co...
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American Society for Microbiology
2019
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oai:doaj.org-article:3cbe7693f30c4d82ba23a68fa6ecdc4b2021-11-15T15:59:41ZA Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the <named-content content-type="genus-species">Plasmodium falciparum</named-content> Life Cycle10.1128/mBio.01170-192150-7511https://doaj.org/article/3cbe7693f30c4d82ba23a68fa6ecdc4b2019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01170-19https://doaj.org/toc/2150-7511ABSTRACT Plasmodium falciparum has a complex life cycle that involves interaction with multiple tissues inside the human and mosquito hosts. Identification of essential genes at all different stages of the P. falciparum life cycle is urgently required for clinical development of tools for malaria control and eradication. However, the study of P. falciparum is limited by the inability to genetically modify the parasite throughout its life cycle with the currently available genetic tools. Here, we describe the detailed characterization of a new marker-free P. falciparum parasite line that expresses rapamycin-inducible Cre recombinase across the full life cycle. Using this parasite line, we were able to conditionally delete the essential invasion ligand AMA1 in three different developmental stages for the first time. We further confirm efficient gene deletion by targeting the nonessential kinase FIKK7.1. IMPORTANCE One of the major limitations in studying P. falciparum is that so far only asexual stages are amenable to rapid conditional genetic modification. The most promising drug targets and vaccine candidates, however, have been refractory to genetic modification because they are essential during the blood stage or for transmission in the mosquito vector. This leaves a major gap in our understanding of parasite proteins in most life cycle stages and hinders genetic validation of drug and vaccine targets. Here, we describe a method that supports conditional gene deletion across the P. falciparum life cycle for the first time. We demonstrate its potential by deleting essential and nonessential genes at different parasite stages, which opens up completely new avenues for the study of malaria and drug development. It may also allow the realization of novel vaccination strategies using attenuated parasites.Marta TibúrcioAnnie S. P. YangKazuhide YahataPablo Suárez-CortésHugo BeldaSebastian BaumgartenMarga van de Vegte-BolmerGeert-Jan van GemertYouri van WaardenburgElena A. LevashinaRobert W. SauerweinMoritz TreeckAmerican Society for MicrobiologyarticlePlasmodium falciparummalariamolecular methodsreverse genetic analysisMicrobiologyQR1-502ENmBio, Vol 10, Iss 5 (2019) |
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Plasmodium falciparum malaria molecular methods reverse genetic analysis Microbiology QR1-502 |
| spellingShingle |
Plasmodium falciparum malaria molecular methods reverse genetic analysis Microbiology QR1-502 Marta Tibúrcio Annie S. P. Yang Kazuhide Yahata Pablo Suárez-Cortés Hugo Belda Sebastian Baumgarten Marga van de Vegte-Bolmer Geert-Jan van Gemert Youri van Waardenburg Elena A. Levashina Robert W. Sauerwein Moritz Treeck A Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the <named-content content-type="genus-species">Plasmodium falciparum</named-content> Life Cycle |
| description |
ABSTRACT Plasmodium falciparum has a complex life cycle that involves interaction with multiple tissues inside the human and mosquito hosts. Identification of essential genes at all different stages of the P. falciparum life cycle is urgently required for clinical development of tools for malaria control and eradication. However, the study of P. falciparum is limited by the inability to genetically modify the parasite throughout its life cycle with the currently available genetic tools. Here, we describe the detailed characterization of a new marker-free P. falciparum parasite line that expresses rapamycin-inducible Cre recombinase across the full life cycle. Using this parasite line, we were able to conditionally delete the essential invasion ligand AMA1 in three different developmental stages for the first time. We further confirm efficient gene deletion by targeting the nonessential kinase FIKK7.1. IMPORTANCE One of the major limitations in studying P. falciparum is that so far only asexual stages are amenable to rapid conditional genetic modification. The most promising drug targets and vaccine candidates, however, have been refractory to genetic modification because they are essential during the blood stage or for transmission in the mosquito vector. This leaves a major gap in our understanding of parasite proteins in most life cycle stages and hinders genetic validation of drug and vaccine targets. Here, we describe a method that supports conditional gene deletion across the P. falciparum life cycle for the first time. We demonstrate its potential by deleting essential and nonessential genes at different parasite stages, which opens up completely new avenues for the study of malaria and drug development. It may also allow the realization of novel vaccination strategies using attenuated parasites. |
| format |
article |
| author |
Marta Tibúrcio Annie S. P. Yang Kazuhide Yahata Pablo Suárez-Cortés Hugo Belda Sebastian Baumgarten Marga van de Vegte-Bolmer Geert-Jan van Gemert Youri van Waardenburg Elena A. Levashina Robert W. Sauerwein Moritz Treeck |
| author_facet |
Marta Tibúrcio Annie S. P. Yang Kazuhide Yahata Pablo Suárez-Cortés Hugo Belda Sebastian Baumgarten Marga van de Vegte-Bolmer Geert-Jan van Gemert Youri van Waardenburg Elena A. Levashina Robert W. Sauerwein Moritz Treeck |
| author_sort |
Marta Tibúrcio |
| title |
A Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the <named-content content-type="genus-species">Plasmodium falciparum</named-content> Life Cycle |
| title_short |
A Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the <named-content content-type="genus-species">Plasmodium falciparum</named-content> Life Cycle |
| title_full |
A Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the <named-content content-type="genus-species">Plasmodium falciparum</named-content> Life Cycle |
| title_fullStr |
A Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the <named-content content-type="genus-species">Plasmodium falciparum</named-content> Life Cycle |
| title_full_unstemmed |
A Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the <named-content content-type="genus-species">Plasmodium falciparum</named-content> Life Cycle |
| title_sort |
novel tool for the generation of conditional knockouts to study gene function across the <named-content content-type="genus-species">plasmodium falciparum</named-content> life cycle |
| publisher |
American Society for Microbiology |
| publishDate |
2019 |
| url |
https://doaj.org/article/3cbe7693f30c4d82ba23a68fa6ecdc4b |
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