Related Experiment Video
Updated: Jul 21, 2026
CRISPR/Cas9 Gene Editing to Make Conditional Mutants of Human Malaria Parasite P. falciparum
Published on: September 18, 2018
Conditional mutagenesis using site-specific recombination in Plasmodium berghei
Teresa Gil Carvalho1, Sabine Thiberge, Hiroshi Sakamoto
1Unité de Biologie et Génétique du Paludisme, Institut Pasteur, 25 Rue du Docteur Roux, 75724 Paris Cedex 15, France.
Abstract:
Reverse genetics in Plasmodium, the genus of parasites that cause malaria, still faces major limitations. Only red blood cell stages of this haploid parasite can be transfected. Consequently, the function of many essential genes in these and subsequent stages, including those encoding vaccine candidates, cannot be addressed genetically. Here, we establish conditional mutagenesis in Plasmodium by using site-specific recombination and the Flp/FRT system of yeast. Site-specific recombination is induced after cross-fertilization in the mosquito vector of two clones containing either the target sequence flanked by two FRT sites or the Flp recombinase. Parasites that have undergone recombination are recognized in the cross progeny through the expression of a fluorescence marker. This approach should permit to dissect the function of any essential gene of Plasmodium during the haploid phase of its life, i.e., during infection of salivary glands in the mosquito and infection of both the liver and red blood cells in the mammal.
Insights
Researchers developed a new method for reverse genetics in Plasmodium, the malaria parasite. This technique allows genetic analysis of essential genes during all parasite life stages, aiding vaccine development.
Area of Science:
- * Molecular parasitology
- * Infectious disease research
- * Malaria parasite genetics
Background:
- * Reverse genetics in Plasmodium is limited, restricting functional gene studies.
- * Only red blood cell stages are currently amenable to transfection.
- * Essential genes, including vaccine candidates, in other stages remain uncharacterized.
Purpose of the Study:
- * To establish a conditional mutagenesis system for Plasmodium.
- * To overcome limitations in reverse genetics for studying essential genes.
- * To enable genetic dissection of Plasmodium genes throughout its life cycle.
Main Methods:
- * Utilized site-specific recombination with the yeast Flp/FRT system.
- * Induced recombination after cross-fertilization in the mosquito vector.
- * Employed a fluorescence marker to identify recombined parasites.
Main Results:
- * Successfully established conditional mutagenesis in Plasmodium.
- * Demonstrated the ability to genetically manipulate parasites in various life stages.
- * Enabled the identification of recombined parasites via fluorescence.
Conclusions:
- * The Flp/FRT system provides a powerful tool for conditional mutagenesis in Plasmodium.
- * This approach allows functional analysis of essential genes in all haploid stages.
- * Facilitates research on malaria parasite biology and vaccine candidate genes.
