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Genomic Transformation of the Picoeukaryote Ostreococcus tauri

Common problems hindering rapid progress in Plant Sciences include cellular, tissue and whole organism complexity, and notably the high level of genomic redundancy affecting simple genetics in higher plants. The novel model organism Ostreococcus tauri is the smallest free-living eukaryote known to d...

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Autores principales: van Ooijen, Gerben, Knox, Kirsten, Kis, Katalin, Bouget, François-Yves, Millar, Andrew J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3476405/
https://www.ncbi.nlm.nih.gov/pubmed/22825291
http://dx.doi.org/10.3791/4074
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author van Ooijen, Gerben
Knox, Kirsten
Kis, Katalin
Bouget, François-Yves
Millar, Andrew J.
author_facet van Ooijen, Gerben
Knox, Kirsten
Kis, Katalin
Bouget, François-Yves
Millar, Andrew J.
author_sort van Ooijen, Gerben
collection PubMed
description Common problems hindering rapid progress in Plant Sciences include cellular, tissue and whole organism complexity, and notably the high level of genomic redundancy affecting simple genetics in higher plants. The novel model organism Ostreococcus tauri is the smallest free-living eukaryote known to date, and possesses a greatly reduced genome size and cellular complexity(1,2), manifested by the presence of just one of most organelles (mitochondrion, chloroplast, golgi stack) per cell, and a genome containing only ~8000 genes. Furthermore, the combination of unicellularity and easy culture provides a platform amenable to chemical biology approaches. Recently, Ostreococcus has been successfully employed to study basic mechanisms underlying circadian timekeeping(3-6). Results from this model organism have impacted not only plant science, but also mammalian biology(7). This example highlights how rapid experimentation in a simple eukaryote from the green lineage can accelerate research in more complex organisms by generating testable hypotheses using methods technically feasible only in this background of reduced complexity. Knowledge of a genome and the possibility to modify genes are essential tools in any model species. Genomic(1), Transcriptomic(8), and Proteomic(9) information for this species is freely available, whereas the previously reported methods(6,10) to genetically transform Ostreococcus are known to few laboratories worldwide. In this article, the experimental methods to genetically transform this novel model organism with an overexpression construct by means of electroporation are outlined in detail, as well as the method of inclusion of transformed cells in low percentage agarose to allow selection of transformed lines originating from a single transformed cell. Following the successful application of Ostreococcus to circadian research, growing interest in Ostreococcus can be expected from diverse research areas within and outside plant sciences, including biotechnological areas. Researchers from a broad range of biological and medical sciences that work on conserved biochemical pathways may consider pursuing research in Ostreococcus, free from the genomic and organismal complexity of larger model species.
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spelling pubmed-34764052012-10-24 Genomic Transformation of the Picoeukaryote Ostreococcus tauri van Ooijen, Gerben Knox, Kirsten Kis, Katalin Bouget, François-Yves Millar, Andrew J. J Vis Exp Microbiology Common problems hindering rapid progress in Plant Sciences include cellular, tissue and whole organism complexity, and notably the high level of genomic redundancy affecting simple genetics in higher plants. The novel model organism Ostreococcus tauri is the smallest free-living eukaryote known to date, and possesses a greatly reduced genome size and cellular complexity(1,2), manifested by the presence of just one of most organelles (mitochondrion, chloroplast, golgi stack) per cell, and a genome containing only ~8000 genes. Furthermore, the combination of unicellularity and easy culture provides a platform amenable to chemical biology approaches. Recently, Ostreococcus has been successfully employed to study basic mechanisms underlying circadian timekeeping(3-6). Results from this model organism have impacted not only plant science, but also mammalian biology(7). This example highlights how rapid experimentation in a simple eukaryote from the green lineage can accelerate research in more complex organisms by generating testable hypotheses using methods technically feasible only in this background of reduced complexity. Knowledge of a genome and the possibility to modify genes are essential tools in any model species. Genomic(1), Transcriptomic(8), and Proteomic(9) information for this species is freely available, whereas the previously reported methods(6,10) to genetically transform Ostreococcus are known to few laboratories worldwide. In this article, the experimental methods to genetically transform this novel model organism with an overexpression construct by means of electroporation are outlined in detail, as well as the method of inclusion of transformed cells in low percentage agarose to allow selection of transformed lines originating from a single transformed cell. Following the successful application of Ostreococcus to circadian research, growing interest in Ostreococcus can be expected from diverse research areas within and outside plant sciences, including biotechnological areas. Researchers from a broad range of biological and medical sciences that work on conserved biochemical pathways may consider pursuing research in Ostreococcus, free from the genomic and organismal complexity of larger model species. MyJove Corporation 2012-07-13 /pmc/articles/PMC3476405/ /pubmed/22825291 http://dx.doi.org/10.3791/4074 Text en Copyright © 2012, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Microbiology
van Ooijen, Gerben
Knox, Kirsten
Kis, Katalin
Bouget, François-Yves
Millar, Andrew J.
Genomic Transformation of the Picoeukaryote Ostreococcus tauri
title Genomic Transformation of the Picoeukaryote Ostreococcus tauri
title_full Genomic Transformation of the Picoeukaryote Ostreococcus tauri
title_fullStr Genomic Transformation of the Picoeukaryote Ostreococcus tauri
title_full_unstemmed Genomic Transformation of the Picoeukaryote Ostreococcus tauri
title_short Genomic Transformation of the Picoeukaryote Ostreococcus tauri
title_sort genomic transformation of the picoeukaryote ostreococcus tauri
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3476405/
https://www.ncbi.nlm.nih.gov/pubmed/22825291
http://dx.doi.org/10.3791/4074
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