ArticlePloS one2017
Plasmid-free CRISPR/Cas9 genome editing in Plasmodium falciparum confirms mutations conferring resistance to the dihydroisoquinolone clinical candidate SJ733.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 56 citations in OpenAlex.
- Acetyl-CoA synthetase mutations affect the susceptibility ofMicrobiology spectrum · 2025Article
- A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.Nucleic acids research · 2025Article
- CRISPR-Cas13 in malaria parasite: Diagnosis and prospective gene function identification.Frontiers in microbiology · 2023Review
- CRISPR-Cas9: Taming protozoan parasites with bacterial scissor.Journal of parasitic diseases : official organ of the Indian Society for Parasitology · 2022Review
- A G358S mutation in the Plasmodium falciparum NaNature communications · 2022Article
- CRISPR/Cas9 Editing of the Plasmodium falciparum Genome.Methods in molecular biology (Clifton, N.J.) · 2022Review
- Associations between Varied Susceptibilities to PfATP4 Inhibitors and Genotypes in Ugandan Plasmodium falciparum Isolates.Antimicrobial agents and chemotherapy · 2021Article
- Highly efficient CRISPR/Cas9 system in Plasmodium falciparum using Cas9-expressing parasites and a linear donor template.Scientific reports · 2021Article
- CRISPR/Cas9 Ribonucleoprotein-Based Genome Editing Methodology in the Marine Protozoan ParasiteFrontiers in bioengineering and biotechnology · 2021Article
- FnCas12a/crRNA-Mediated Genome Editing inFrontiers in genetics · 2021Article
- Recent Progress in the Development of New Antimalarial Drugs with Novel Targets.Drug design, development and therapy · 2020Review
- The resistome and genomic reconnaissance in the age of malaria elimination.Disease models & mechanisms · 2019Review
- Cutting back malaria: CRISPR/Cas9 genome editing of Plasmodium.Briefings in functional genomics · 2019Review
- Generation and functional characterisation of Plasmodium yoelii csp deletion mutants using a microhomology-based CRISPR/Cas9 method.International journal for parasitology · 2019Article
- Article
- Ribozyme-mediated, multiplex CRISPR gene editing and CRISPR interference (CRISPRi) in rodent-infectiousThe Journal of biological chemistry · 2019Article
- Plasmodium falciparum In Vitro Drug Resistance Selections and Gene Editing.Methods in molecular biology (Clifton, N.J.) · 2019Article
- CRISPR/Cas9 Gene Editing to Make Conditional Mutants of Human Malaria Parasite P. falciparum.Journal of visualized experiments : JoVE · 2018Article
- New and emerging uses of CRISPR/Cas9 to genetically manipulate apicomplexan parasites.Parasitology · 2018Review
- A Cas9 transgenic Plasmodium yoelii parasite for efficient gene editing.Molecular and biochemical parasitology · 2018Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Genetic manipulation of the deadly malaria parasite Plasmodium falciparum remains challenging, but the rise of CRISPR/Cas9-based genome editing tools is increasing the feasibility of altering this parasite's genome in order to study its biology. Of particular interest is the investigation of drug targets and drug resistance mechanisms, which have major implications for fighting malaria. We present a new method for introducing drug resistance mutations in P. falciparum without the use of plasmids or the need for cloning homologous recombination templates. We demonstrate this method by introducing edits into the sodium efflux channel PfATP4 by transfection of a purified CRISPR/Cas9-guide RNA ribonucleoprotein complex and a 200-nucleotide single-stranded oligodeoxynucleotide (ssODN) repair template. Analysis of whole genome sequencing data with the variant-finding program MinorityReport confirmed that only the intended edits were made, and growth inhibition assays confirmed that these mutations confer resistance to the antimalarial SJ733. The method described here is ideally suited for the introduction of mutations that confer a fitness advantage under selection conditions, and the novel finding that an ssODN can function as a repair template in P. falciparum could greatly simplify future editing attempts regardless of the nuclease used or the delivery method.
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