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Biolistic Transformation of Chlamydomonas reinhardtii and Saccharomyces cerevisiae Mitochondria

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Mitochondrial DNA

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2615))

Abstract

Chlamydomonas reinhardtii and Saccharomyces cerevisiae are currently the two micro-organisms in which genetic transformation of mitochondria is routinely performed. The generation of a large variety of defined alterations as well as the insertion of ectopic genes in the mitochondrial genome (mtDNA) are possible, especially in yeast. Biolistic transformation of mitochondria is achieved through the bombardment of microprojectiles coated with DNA, which can be incorporated into mtDNA thanks to the highly efficient homologous recombination machinery present in S. cerevisiae and C. reinhardtii organelles. Despite a low frequency of transformation, the isolation of transformants in yeast is relatively quick and easy, since several natural or artificial selectable markers are available, while the selection in C. reinhardtii remains long and awaits new markers. Here, we describe the materials and techniques used to perform biolistic transformation, in order to mutagenize endogenous mitochondrial genes or insert novel markers into mtDNA. Although alternative strategies to edit mtDNA are being set up, so far, insertion of ectopic genes relies on the biolistic transformation techniques.

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Acknowledgments

We are indebted to Claudia Serot for generating the supplementary video showing the bombardment procedure, including filming, choosing the sequences and editing the video clip. N. B. is supported by the Centre National pour la Recherche Scientifique, and C.R. is supported by Fonds National de la Recherche scientifique (FNRS, CDR J.0175.20) and Fonds Wetenschappelijk Onderzoek—Vlaanderen (FWO) and FNRS under EOS Project No. 30829584.

Author information

Authors and Affiliations

  1. Institute of Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette cedex, France

    Nathalie Bonnefoy

  2. Genetics and physiology of microalgae, InBios/Phytosystems Research Unit, University of Liege, Liege, Belgium

    Claire Remacle

Authors
  1. Nathalie Bonnefoy
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  2. Claire Remacle
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Corresponding author

Correspondence to Claire Remacle .

Editor information

Editors and Affiliations

  1. Wellcome Centre for Mitochondrial Research, Newcastle University, Newcastle upon Tyne, UK

    Thomas J. Nicholls

  2. Department of Medical Biochemistry and Cell Biology, University of Gothenburg, Gothenburg, Sweden

    Jay P. Uhler

  3. Department of Medical Biochemistry and Cell Biology, University of Gothenburg, Gothenburg, Sweden

    Maria Falkenberg

1 Electronic Supplementary Material

Example of bombardment procedure (MP4 86151 kb)

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Bonnefoy, N., Remacle, C. (2023). Biolistic Transformation of Chlamydomonas reinhardtii and Saccharomyces cerevisiae Mitochondria. In: Nicholls, T.J., Uhler, J.P., Falkenberg, M. (eds) Mitochondrial DNA. Methods in Molecular Biology, vol 2615. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2922-2_24

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  • DOI: https://doi.org/10.1007/978-1-0716-2922-2_24

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2921-5

  • Online ISBN: 978-1-0716-2922-2

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