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Updated: Oct 20, 2025
CRISPR/Cas9 Gene Editing to Make Conditional Mutants of Human Malaria Parasite P. falciparum
Published on: September 18, 2018
Genetic Manipulation of Non-Falciparum Human Malaria Parasites
Taís Baruel Vieira1, Thafne Plastina Astro1, Roberto Rudge de Moraes Barros1
1Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.
Abstract:
The development of genetic manipulation of Plasmodium falciparum in the 1980s was key to study malaria biology. Genetically modified parasites have been used to study several aspects of the disease, such as red blood cell invasion, drug resistance mechanisms, gametocyte development and mosquito transmission. However, biological and genetic differences between P. falciparum and the other human malaria parasites make P. falciparum a poor model to study different species. The lack of robust systems of long-term in vitro culture of P. vivax and the other human malaria parasites lagged the genetic manipulation of these species. Here we review the efforts to generate genetically modified non-falciparum human malaria parasites, in vivo and in vitro. Using in vivo models - infection of non-human primates such as rhesus macaques and saimiri monkeys - researchers were able to generate transgenic lines of P. knowlesi, P. cynomolgi, and P. vivax. The development of long-term in vitro culture of P. knowlesi in the 2000's, using rhesus and human red blood cells, created a platform to genetically manipulate non-falciparum malaria parasites. Recently, the use of CRISPR/Cas9 technology to genome edit P. knowlesi provides another tool to non-falciparum malaria research, extending the possibilities and allowing researchers to study different aspects of the biology of these parasites and understand the differences between these species and P. falciparum.
Insights
Genetic tools now enable the study of non-falciparum malaria parasites like Plasmodium vivax. Advances in in vitro culture and CRISPR technology are crucial for understanding malaria parasite biology beyond Plasmodium falciparum.
Area of Science:
- Malariology
- Parasitology
- Genetics
Background:
- Genetic manipulation of Plasmodium falciparum since the 1980s has been vital for malaria research.
- Differences between Plasmodium falciparum and other human malaria parasites limit its use as a model for diverse species.
- Challenges in long-term in vitro culture of Plasmodium vivax and other non-falciparum species have hindered genetic studies.
Purpose of the Study:
- To review efforts in generating genetically modified non-falciparum human malaria parasites.
- To highlight advancements in both in vivo and in vitro genetic manipulation techniques for these parasites.
- To discuss the implications of these advancements for understanding malaria biology and inter-species differences.
Main Methods:
- Utilizing in vivo models, including non-human primates (rhesus macaques, saimiri monkeys), to generate transgenic lines.
- Developing robust long-term in vitro culture systems for Plasmodium knowlesi using rhesus and human red blood cells.
- Employing CRISPR/Cas9 technology for genome editing in Plasmodium knowlesi.
Main Results:
- Successful generation of transgenic lines for Plasmodium knowlesi, Plasmodium cynomolgi, and Plasmodium vivax using in vivo models.
- Establishment of a platform for genetic manipulation of non-falciparum malaria parasites through improved in vitro culture of P. knowlesi.
- Demonstration of CRISPR/Cas9 as a powerful tool for genome editing in P. knowlesi, expanding research capabilities.
Conclusions:
- Genetic manipulation of non-falciparum malaria parasites is now feasible through in vivo and in vitro strategies.
- Advances in culture and genome editing technologies are critical for studying Plasmodium vivax and other species.
- These tools will enhance our understanding of malaria parasite biology and the distinctions between different Plasmodium species.
