Optimizing CRISPR-Cas12-based detection methods for drug-resistant Plasmodium falciparum with an advanced gene visualization pipeline

Year 2025, Volume: 29 Issue: 3, 1078 - 1088, 04.06.2025

Arli Aditya Parikesit1 Rio Hermantara Elizabeth Sidhartha Daniel Ryan Fugaha Devita Mayanda Heerlie Solmaz Aslanzadeh

https://doi.org/10.12991/jrespharm.1694230

Abstract

Malaria, a life-threatening disease caused by the parasite Plasmodium falciparum, poses a significant health challenge, particularly in tropical regions where it is most prevalent. This disease is traditionally treated with various medications, including chloroquine, quinine, artemisinin, sulfadoxine-pyrimethamine, and combinations of artemisinin-based therapies. However, the emergence of drug-resistant strains of P. falciparum has necessitated the development of more sophisticated diagnostic and treatment strategies to effectively manage and combat this disease. In response to this growing concern, our research project endeavors to pioneer a novel approach to malaria diagnosis and treatment. We focus on the design of Loop-Mediated Isothermal Amplification (LAMP) primers, which are engineered to specifically identify treatment-resistant strains of P. falciparum. The project strongly emphasizes optimizing the guide RNA (gRNA) efficacy, which is pivotal for enhancing the specificity and sensitivity of the CRISPR-Cas12 system in detecting these resistant strains. Our aim is to significantly improve the efficiency and accuracy of malaria treatments, particularly in areas heavily impacted by drug resistance. By leveraging the advancements in CRISPR technology, we anticipate creating a robust tool for the on-field diagnosis of drug-resistant Malaria based on CHOPCHOP and E-CRISPR in silico gene visualization tools. This research addresses a critical gap in the current treatment paradigm and contributes to the broader initiative of employing innovative genetic technologies for public health. Ultimately, the successful implementation of this project could lead to more effective malaria control and treatment strategies, reducing the global burden of this devastating disease.

Keywords

Malaria , CRISPR , LAMP , Cas12 , gRNA , Plasmodium

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