CRISPR/Cas9 gene editing to make conditional mutants of human malaria parasite p. Falciparum

Heather M. Kudyba, David W. Cobb, Anat Florentin, Michelle Krakowiak, Vasant Muralidharan*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


Malaria is a significant cause of morbidity and mortality worldwide. This disease, which primarily affects those living in tropical and subtropical regions, is caused by infection with Plasmodium parasites. The development of more effective drugs to combat malaria can be accelerated by improving our understanding of the biology of this complex parasite. Genetic manipulation of these parasites is key to understanding their biology; however, historically the genome of P. falciparum has been difficult to manipulate. Recently, CRISPR/Cas9 genome editing has been utilized in malaria parasites, allowing for easier protein tagging, generation of conditional protein knockdowns, and deletion of genes. CRISPR/ Cas9 genome editing has proven to be a powerful tool for advancing the field of malaria research. Here, we describe a CRISPR/Cas9 method for generating glmS-based conditional knockdown mutants in P. falciparum. This method is highly adaptable to other types of genetic manipulations, including protein tagging and gene knockouts.

Original languageAmerican English
Article numbere57747
JournalJournal of Visualized Experiments
Issue number139
StatePublished - 18 Sep 2018
Externally publishedYes

Bibliographical note

Funding Information:
We thank Muthugapatti Kandasamy at the University of Georgia (UGA) Biomedical Microscopy Core for technical assistance and Jose-Juan Lopez-Rubio for sharing the pUF1-Cas9 and pL6 plasmids. This work was supported by ARCS Foundation awards to D.W.C. and to H.M.K., UGA startup funds to V.M., grants from the March of Dimes Foundation (Basil O'Connor Starter Scholar Research Award) to V.M., and US National Institutes of Health grants (R00AI099156 and R01AI130139) to V.M. and (T32AI060546) to H.M.K.

Publisher Copyright:
© 2018, Journal of Visualized Experiments. All rights reserved.


  • Genetics
  • Genetics
  • Glms
  • Issue 139
  • Knockdown
  • Malaria
  • Plasmodium


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