Genome editing in the human malaria parasite Plasmodium falciparum using the CRISPR-Cas9 system (original) (raw)
- Brief Communication
- Published: 01 June 2014
- Molly Gorman1,2,
- Cameron Ross Macpherson1,2,
- Rafael Miyazawa Martins1,2,
- Artur Scherf1,2 &
- …
- Jose-Juan Lopez-Rubio1,2 nAff3
Nature Biotechnology volume 32, pages 819–821 (2014)Cite this article
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Abstract
Genome manipulation in the malaria parasite Plasmodium falciparum remains largely intractable and improved genomic tools are needed to further understand pathogenesis and drug resistance. We demonstrated the CRISPR-Cas9 system for use in P. falciparum by disrupting chromosomal loci and generating marker-free, single-nucleotide substitutions with high efficiency. Additionally, an artemisinin-resistant strain was generated by introducing a previously implicated polymorphism, thus illustrating the value of efficient genome editing in malaria research.
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Acknowledgements
We thank L. Mancio-Silva (Instituto de Medicina Molecular, Lisboa) and N. Siegel (Institute for Molecular Infection Biology, Wuerzburg) for critically reading the manuscript and PlasmoDB for the invaluable malaria-database support. F. Zhang laboratory (MIT, Boston) for deposition of pX330 in Addgene. A.B. Vaidya (Drexel University, Philadelphia) for pUF1 plasmid. D. Fidock laboratory (Columbia University, New York) for NF54EGFP strain. C. Buchrieser (Institut Pasteur, Paris) for lending Amaxa 4D electroporator (Lonza) and P. Escoll Guerrero (Institut Pasteur, Paris) for assistance. A. Nacer (Institut Pasteur, Paris) for help with immunofluorescence experiments. GenoScreen team, especially H. Blanquart and A. Gourbeyre, for sequencing. This work was supported by the Agence Nationale de la Recherche (ANR 11 JSV3 004 01 PlasmoPiggyBac), ERC Advanced Grant (PlasmoEscape 250320) and the French Parasitology consortium ParaFrap (ANR-11-LABX0024). Me.G. and Mo.G. were funded by ANR 11 JSV3 004 01 and J.-J.L.-R. by the Institut National de la Santé et de la Recherche Médicale (INSERM).
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- Mehdi Ghorbal & Jose-Juan Lopez-Rubio
Present address: Present address: CNRS 5290/IRD 224/University Montpellier 1&2 (“MiVEGEC”), Montpellier, France.,
Authors and Affiliations
- Biology of Host-Parasite Interactions Unit, Institut Pasteur, Paris, France
Mehdi Ghorbal, Molly Gorman, Cameron Ross Macpherson, Rafael Miyazawa Martins, Artur Scherf & Jose-Juan Lopez-Rubio - CNRS URA 2581, Institut Pasteur, Paris, France
Mehdi Ghorbal, Molly Gorman, Cameron Ross Macpherson, Rafael Miyazawa Martins, Artur Scherf & Jose-Juan Lopez-Rubio
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- Mehdi Ghorbal
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Contributions
J.-J.L.-R. devised the research experimental design with contributions from Me.G. Mo.G., Me.G. and J.-J.L.-R. performed the experiments. R.M.M. prepared the libraries for next-generation sequencing. C.R.M. conducted the bioinformatic analysis. A.S. provided funding. J.-J.L.-R. and Me.G. wrote the manuscript with contributions from all the authors.
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Correspondence toJose-Juan Lopez-Rubio.
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The authors declare no competing financial interests.
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Ghorbal, M., Gorman, M., Macpherson, C. et al. Genome editing in the human malaria parasite Plasmodium falciparum using the CRISPR-Cas9 system.Nat Biotechnol 32, 819–821 (2014). https://doi.org/10.1038/nbt.2925
- Received: 18 February 2014
- Accepted: 08 May 2014
- Published: 01 June 2014
- Issue Date: August 2014
- DOI: https://doi.org/10.1038/nbt.2925