Optimization of CRISPR/Cas System for Malaria Parasites

Malaria is primarily considered as an infectious and life-threatening disease, causing fever, chills, lethargy, nausea, vomiting, diarrhea and other flu-like symptoms in humans. Female Anopheles mosquitos are merely extremely competent vectors of the Plasmodium parasite, the true perpetrator of this disease. There are five species of this unicellular protozoan parasite (P. falciparum, P. vivax, P. ovale, P. knowlesi and P. malariae) that cause malaria in humans. P. falciparum, however, is the most infectious and responsible for severe forms of the disease. According to the latest world malaria report, a total of 229 million cases and 409, 000 malaria deaths were estimated in 2019 (WHO, 2020). 

Recently, CRISPR/Cas9 system, as a versatile gene-editing platform facilitating researches of multiple organisms, has been adopted for genome editing in malaria parasites. However, typical genome editing in the pathogen mediated by CRISPR/Cas9 system demands co-transfection of two episomal plasmids respectively carrying Cas9 endonuclease and a targeting single guide RNA, which results in poor efficiency of gene editing. 

Now, a new study has found a way to circumvent this acute problem. The study, published in the journal Frontiers in Microbiology, shows that an integrative CRISPR/Cas9 system, named CRISPR/Cas9i, can speed up the generation of transgenic strains in P. falciparum and achieve multiplexed genome editing by a single-round transfection in the specie. The new findings may help understand the genetics of malaria parasites and provide alternative tools to facilitate the utilization of Cas9 in malaria research.


                                 Diagram illustrating the Cas9e and Cas9i system (Yuemeng et al., 2021).

The latest work in malaria comes from researchers of Tongji University and our cloud scientist Feng Ding of Wenzhou Medical University. The investigators were curious about whether such an optimized system (Cas9i) could outperform the traditional episomal CRISPR/Cas9 system (Cas9e for short) in P. falciparum. They generated a parental Cas9i line through a single-crossover recombination event, integrating the Cas9 expression cassette into genomic locus and randomly selected several genes for editing by either Cas9i or Cas9e systems. The results showed that the maximum and average time for detecting first resistant parasites were significantly shorter in Cas9i system than that in Cas9e system, indicating an improved genome editing efficiency with the optimized tool. 

Reference
Zhao, Yuemeng, et al. "Optimization of CRISPR/Cas System for Improving Genome Editing Efficiency in Plasmodium falciparum ." Frontiers in Microbiology 11 (2021): 3377.

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Infecting over 200 million people annually, malaria has been around for centuries. See how much you know and what you can learn about the disease through the reward-wining quiz below. A perfect score will bring you a chance to win $20!

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Previous Newsletters

MolecularCloud April Newsletter: How to Rapidly Develop ADC for Cancer Treatments?
MolecularCloud March Newsletter:How Nanotechnology Helped Create mRNA COVID-19 Vaccines?MolecularCloud February Newsletter: Plant Genome Editing by SpCas9 Variants


Q1: Which of the following is not a symptom of Malaria?

  • Hearing loss
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  • Sweating
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  • Fever
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  • Headache
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Q2: The infectious agent that causes malaria is known as which of the following?

  • Protozoan parasite
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  • Bacterial parasite
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  • Viral parasite
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  • Fungal parasite
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Q3: True or false, malaria can not be spread by male mosquito?

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Q4: Malaria parasites can be found worldwide except which of the following?

  • Antarctica
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  • Asia
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  • America
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Q5: Who won a Nobel Prize in Medicine or Physiology for discovering antimalarial compound, artemisinin?

  • Rober Koch
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  • Paul Ehrlich
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  • Youyou Tu
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  • Ronald Ross
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10 Reply

Very Nice quizz. Malaria is a terrible diasease, Id love to see new vaccines and treatments to erradicate It, especially since I'm from Brazil, a country that IS largely affected by Malaria.


This is interesting, I am a Ph.D. student and would like to collaborate with anyone who has funding using CRISPR


Years of research has been directed to developing a vaccine against Plasmodium falciparum with little success. Finally a new approach to tackle this important disease with such a complex life-cycle.


I agree with you, I am a Ph.D. student and will like to collaborate to work on a topic in malaria relating to host genetics and clinical malaria. Let me know if we can talk about this. sdhikrullahi@gmail.com

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