Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

Genome editing in the human malaria parasite Plasmodium falciparum using the CRISPR-Cas9 system

Nature Biotechnology volume 32, pages 819–821 (2014)Cite this article

Subjects

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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Targeted P. falciparum genome editing using sgRNA:Cas9.
Figure 2: Marker-free nucleotide editing using sgRNA:Cas9 in P. falciparum.

Similar content being viewed by others

Accession codes

Primary accessions

BioProject

References

  1. Straimer, J. et al. Nat. Methods 9, 993–998 (2012).

    Article  CAS  Google Scholar 

  2. Bhaya, D., Davison, M. & Barrangou, R. Annu. Rev. Genet. 45, 273–297 (2011).

    Article  CAS  Google Scholar 

  3. Jinek, M. et al. Science 337, 816–821 (2012).

    Article  CAS  Google Scholar 

  4. Kirkman, L.A., Lawrence, E.A. & Deitsch, K.W. Nucleic Acids Res. 42, 370–379 (2014).

    Article  CAS  Google Scholar 

  5. Cong, L. et al. Science 339, 819–823 (2013).

    Article  CAS  Google Scholar 

  6. Bawankar, P., Shaw, P.J., Sardana, R., Babar, P.H. & Patankar, S. Mol. Biol. Rep. 37, 2125–2133 (2010).

    Article  CAS  Google Scholar 

  7. Duraisingh, M.T., Triglia, T. & Cowman, A.F. Int. J. Parasitol. 32, 81–89 (2002).

    Article  CAS  Google Scholar 

  8. Deitsch, K., Driskill, C. & Wellems, T. Nucleic Acids Res. 29, 850–853 (2001).

    Article  CAS  Google Scholar 

  9. Pfander, C. et al. Nat. Methods 8, 1078–1082 (2011).

    Article  CAS  Google Scholar 

  10. Crabb, B.S. et al. Cell 89, 287–296 (1997).

    Article  CAS  Google Scholar 

  11. Moon, R.W. et al. Proc. Natl. Acad. Sci. USA 110, 531–536 (2013).

    Article  CAS  Google Scholar 

  12. Deshmukh, A.S. et al. Nucleic Acids Res. 40, 5313–5331 (2012).

    Article  CAS  Google Scholar 

  13. Ariey, F. et al. Nature 505, 50–55 (2014).

    Article  Google Scholar 

  14. Ran, F.A. et al. Nat. Protoc. 8, 2281–2308 (2013).

    Article  CAS  Google Scholar 

  15. Gilbert, L.A. et al. Cell 154, 442–451 (2013).

    Article  CAS  Google Scholar 

  16. Ganesan, S.M. et al. Mol. Biochem. Parasitol. 177, 29–34 (2011).

    Article  CAS  Google Scholar 

  17. Mali, P. et al. Science 339, 823–826 (2013).

    Article  CAS  Google Scholar 

  18. Wu, Y., Sifri, C.D., Lei, H.H., Su, X.Z. & Wellems, T.E. Proc. Natl. Acad. Sci. USA 92, 973–977 (1995).

    Article  CAS  Google Scholar 

  19. Collins, C.R. et al. Mol. Microbiol. 88, 687–701 (2013).

    Article  CAS  Google Scholar 

  20. Salanti, A. et al. Mol. Microbiol. 49, 179–191 (2003).

    Article  CAS  Google Scholar 

  21. Mancio-Silva, L., Rojas-Meza, A.P., Vargas, M., Scherf, A. & Hernandez-Rivas, R. J. Cell Sci. 121, 2046–2053 (2008).

    Article  CAS  Google Scholar 

  22. Witkowski, B. et al. Lancet Infect. Dis. 13, 1043–1049 (2013).

    Article  CAS  Google Scholar 

Download references

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).

Author information

Author notes

  1. Mehdi Ghorbal & Jose-Juan Lopez-Rubio

    Present address: Present address: CNRS 5290/IRD 224/University Montpellier 1&2 (“MiVEGEC”), Montpellier, France.,

Authors and Affiliations

  1. 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

  2. CNRS URA 2581, Institut Pasteur, Paris, France

    Mehdi Ghorbal, Molly Gorman, Cameron Ross Macpherson, Rafael Miyazawa Martins, Artur Scherf & Jose-Juan Lopez-Rubio

Authors
  1. Mehdi Ghorbal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Molly Gorman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Cameron Ross Macpherson
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rafael Miyazawa Martins
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Artur Scherf
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jose-Juan Lopez-Rubio
    View author publications

    You can also search for this author in PubMed Google Scholar

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.

Corresponding author

Correspondence to Jose-Juan Lopez-Rubio.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Tables 1 and 2, and Supplementary Results (PDF 1964 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt.2925

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing