ArticleParasites & vectors2017
Tagging to endogenous genes of Plasmodium falciparum using CRISPR/Cas9.
Article in Parasites & vectors, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 34 citations in OpenAlex.
- A heterologous marker-free selection approach for CRISPR/Cas9-based gene editing in the malaria parasitemSphere · 2026Article
- Impact of Fluorophore and Epitope Position on Destabilized Reporter Performance inbioRxiv : the preprint server for biology · 2026Article
- Acetyl-CoA synthetase mutations affect the susceptibility ofMicrobiology spectrum · 2025Article
- CRISPR-Cas Technology as a Revolutionary Genome Editing tool: Mechanisms and Biomedical Applications.Iranian biomedical journal · 2023Review
- Paving the Way: Contributions of Big Data to Apicomplexan and Kinetoplastid Research.Frontiers in cellular and infection microbiology · 2022Review
- Efficient generation of mNeonGreenFrontiers in cellular and infection microbiology · 2022Article
- Optimization of CRISPR/Cas System for Improving Genome Editing Efficiency inFrontiers in microbiology · 2020Article
- Cutting back malaria: CRISPR/Cas9 genome editing of Plasmodium.Briefings in functional genomics · 2019Review
- ABiology open · 2019Article
- Plasmodium falciparum In Vitro Drug Resistance Selections and Gene Editing.Methods in molecular biology (Clifton, N.J.) · 2019Article
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Authors and funding
9 authors at 4 institutions in 1 country.
Funding
Abstract
backgroundPlasmodium falciparum is the deadliest malaria parasite. Currently, there are seldom commercial antibodies against P. falciparum proteins, which greatly limits the study on Plasmodium. CRISPR/Cas9 is an efficient genome editing method, which has been employed in various organisms. However, the use of this technique in P. falciparum is still limited to gene knockout, site-specific mutation and generation of green fluorescent protein (GFP) reporter line with disruption of inserted sites.
resultsWe have adapted the CRISPR/Cas9 system to add commercial tag sequences to endogenous genes of P. falciparum. To add HA or HA-TY1 tags to ck2β1, ck2α and stk, pL6cs-hDHFR-ck2β1/ck2α/stk was constructed, which contained sequences of tags, specific homologous arms, and sgRNA. The P. falciparum 3D7 strain was subsequently transfected with pUF1-BSD-Cas9 and pL6cs-hDHFR-ck2β1/ck2α/stk plasmids via electroporation. After that, BSD and WR99210 drugs were added to the culture to screen parasites containing both plasmids. Twenty days after electroporation, live parasites appeared and were collected to check the tagging by PCR, DNA sequencing, Western blotting and immuno-fluorescence assays. The results showed that the tags were successfully integrated into the C-terminus of these three proteins.
conclusionsWe have improved the method to integrate tags to Plasmodium falciparum genes using the CRISPR/Cas9 method, which lays the foundation for further study of Plasmodium falciparum at the molecular level.
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