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High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery

BACKGROUND: Malaria, a major public health issue in developing nations, is responsible for more than one million deaths a year. The most lethal species, Plasmodium falciparum, causes up to 90% of fatalities. Drug resistant strains to common therapies have emerged worldwide and recent artemisinin-bas...

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Autores principales: Cervantes, Serena, Prudhomme, Jacques, Carter, David, Gopi, Krishna G, Li, Qian, Chang, Young-Tae, Le Roch, Karine G
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2702272/
https://www.ncbi.nlm.nih.gov/pubmed/19515257
http://dx.doi.org/10.1186/1471-2121-10-45
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author Cervantes, Serena
Prudhomme, Jacques
Carter, David
Gopi, Krishna G
Li, Qian
Chang, Young-Tae
Le Roch, Karine G
author_facet Cervantes, Serena
Prudhomme, Jacques
Carter, David
Gopi, Krishna G
Li, Qian
Chang, Young-Tae
Le Roch, Karine G
author_sort Cervantes, Serena
collection PubMed
description BACKGROUND: Malaria, a major public health issue in developing nations, is responsible for more than one million deaths a year. The most lethal species, Plasmodium falciparum, causes up to 90% of fatalities. Drug resistant strains to common therapies have emerged worldwide and recent artemisinin-based combination therapy failures hasten the need for new antimalarial drugs. Discovering novel compounds to be used as antimalarials is expedited by the use of a high-throughput screen (HTS) to detect parasite growth and proliferation. Fluorescent dyes that bind to DNA have replaced expensive traditional radioisotope incorporation for HTS growth assays, but do not give additional information regarding the parasite stage affected by the drug and a better indication of the drug's mode of action. Live cell imaging with RNA dyes, which correlates with cell growth and proliferation, has been limited by the availability of successful commercial dyes. RESULTS: After screening a library of newly synthesized stryrl dyes, we discovered three RNA binding dyes that provide morphological details of live parasites. Utilizing an inverted confocal imaging platform, live cell imaging of parasites increases parasite detection, improves the spatial and temporal resolution of the parasite under drug treatments, and can resolve morphological changes in individual cells. CONCLUSION: This simple one-step technique is suitable for automation in a microplate format for novel antimalarial compound HTS. We have developed a new P. falciparum RNA high-content imaging growth inhibition assay that is robust with time and energy efficiency.
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spelling pubmed-27022722009-06-27 High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery Cervantes, Serena Prudhomme, Jacques Carter, David Gopi, Krishna G Li, Qian Chang, Young-Tae Le Roch, Karine G BMC Cell Biol Methodology Article BACKGROUND: Malaria, a major public health issue in developing nations, is responsible for more than one million deaths a year. The most lethal species, Plasmodium falciparum, causes up to 90% of fatalities. Drug resistant strains to common therapies have emerged worldwide and recent artemisinin-based combination therapy failures hasten the need for new antimalarial drugs. Discovering novel compounds to be used as antimalarials is expedited by the use of a high-throughput screen (HTS) to detect parasite growth and proliferation. Fluorescent dyes that bind to DNA have replaced expensive traditional radioisotope incorporation for HTS growth assays, but do not give additional information regarding the parasite stage affected by the drug and a better indication of the drug's mode of action. Live cell imaging with RNA dyes, which correlates with cell growth and proliferation, has been limited by the availability of successful commercial dyes. RESULTS: After screening a library of newly synthesized stryrl dyes, we discovered three RNA binding dyes that provide morphological details of live parasites. Utilizing an inverted confocal imaging platform, live cell imaging of parasites increases parasite detection, improves the spatial and temporal resolution of the parasite under drug treatments, and can resolve morphological changes in individual cells. CONCLUSION: This simple one-step technique is suitable for automation in a microplate format for novel antimalarial compound HTS. We have developed a new P. falciparum RNA high-content imaging growth inhibition assay that is robust with time and energy efficiency. BioMed Central 2009-06-10 /pmc/articles/PMC2702272/ /pubmed/19515257 http://dx.doi.org/10.1186/1471-2121-10-45 Text en Copyright © 2009 Cervantes et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Article
Cervantes, Serena
Prudhomme, Jacques
Carter, David
Gopi, Krishna G
Li, Qian
Chang, Young-Tae
Le Roch, Karine G
High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery
title High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery
title_full High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery
title_fullStr High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery
title_full_unstemmed High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery
title_short High-content live cell imaging with RNA probes: advancements in high-throughput antimalarial drug discovery
title_sort high-content live cell imaging with rna probes: advancements in high-throughput antimalarial drug discovery
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2702272/
https://www.ncbi.nlm.nih.gov/pubmed/19515257
http://dx.doi.org/10.1186/1471-2121-10-45
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