Abstract
In Plasmodium falciparum, the parasite responsible for the most severe forms of human malaria, many fundamental processes are controlled at the transcriptional level. Studies on diverse aspects of basic parasite biology as well as molecular epidemiology studies often rely on the ability to accurately measure transcript levels, but this is complicated by the cyclic expression patterns of the majority of malaria parasite genes. Here, we provide a complete workflow to measure transcript levels in P. falciparum intraerythrocytic blood stages, overcoming the confounding factors that are commonly encountered. The method described covers all the steps from synchronization of parasite cultures to reverse transcriptase quantitative PCR (RT-qPCR) analysis.
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Acknowledgments
The team is funded by grants from the Spanish Ministry of Science and Innovation (MICINN)/ Agencia Estatal de Investigación (AEI) [SAF2016-76190-R and PID2019-107232RB-I00 to A.C.], co-funded by the European Regional Development Fund (ERDF, European Union), and from “la Caixa” Banking Foundation under the project code HR18-00267 (awarded to A.C.). H.P.P. is funded by the TransGlobalHealth–Erasmus Mundus Joint Doctorate Programme, European Union (scholarship number 2016-1346). A.K.P. is supported by a fellowship from the Secretary for Universities and Research, Catalan Government (FI_B 00373), co-funded by the European Social Fund (ESF), European Commission. Our research is part of ISGlobal’s Program on the Molecular Mechanisms of Malaria, which is partially supported by the Fundación Ramón Areces. We acknowledge support from the Spanish Ministry of Science and Innovation through the “Centro de Excelencia Severo Ochoa 2019-2023” Program (CEX2018-000806-S), and support from the Generalitat de Catalunya through the CERCA Program. Núria Casas-Vila and Anastasia K. Pickford contributed equally to this work.
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Casas-Vila, N., Pickford, A.K., Portugaliza, H.P., Tintó-Font, E., Cortés, A. (2021). Transcriptional Analysis of Tightly Synchronized Plasmodium falciparum Intraerythrocytic Stages by RT-qPCR. In: de Pablos, L.M., Sotillo, J. (eds) Parasite Genomics. Methods in Molecular Biology, vol 2369. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1681-9_10
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DOI: https://doi.org/10.1007/978-1-0716-1681-9_10
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