ArticleNature protocols2011
FLP/FRT-mediated conditional mutagenesis in pre-erythrocytic stages of Plasmodium berghei.
Article in Nature protocols, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 63 citations in OpenAlex.
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- Novel systems to study vector-pathogen interactions in malaria.Frontiers in cellular and infection microbiology · 2023Review
- A toolbox for conditional control of gene expression in apicomplexan parasites.Molecular microbiology · 2022Review
- An essential vesicular-trafficking phospholipase mediates neutral lipid synthesis and contributes to hemozoin formation in Plasmodium falciparum.BMC biology · 2021Article
- Conditional Gene Deletion in Mammalian and Mosquito Stages of Plasmodium berghei Using Dimerizable Cre Recombinase.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Plasmodium translocon component EXP2 facilitates hepatocyte invasion.Nature communications · 2020Article
- Lineage tracing: technology tool for exploring the development, regeneration, and disease of the digestive system.Stem cell research & therapy · 2020Review
- Overlapping and distinct roles of CDPK family members in the pre-erythrocytic stages of the rodent malaria parasite, Plasmodium berghei.PLoS pathogens · 2020Article
- The Bradyzoite: A Key Developmental Stage for the Persistence and Pathogenesis of Toxoplasmosis.Pathogens (Basel, Switzerland) · 2020Review
- HIV‑1 integrase inhibitors targeting various DDE transposases: Retroviral integration versus RAG‑mediated recombination (Review).Molecular medicine reports · 2019Review
- Ribozyme-mediated, multiplex CRISPR gene editing and CRISPR interference (CRISPRi) in rodent-infectiousThe Journal of biological chemistry · 2019Article
- PKAc is not required for the preerythrocytic stages ofLife science alliance · 2019Article
- Three classes of response elements for human PRC2 and MLL1/2-Trithorax complexes.Nucleic acids research · 2018Article
- Generation of transgenic rodent malaria parasites by transfection of cell culture-derived merozoites.Malaria journal · 2017Article
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- Plasmodium Merozoite TRAP Family Protein Is Essential for Vacuole Membrane Disruption and Gamete Egress from Erythrocytes.Cell host & microbe · 2016Article
- Invasion of hepatocytes by Plasmodium sporozoites requires cGMP-dependent protein kinase and calcium dependent protein kinase 4.Molecular microbiology · 2016Article
- The IntXO-PSL Recombination System Is a Key Component of the Second Maintenance System for Bacillus anthracis Plasmid pXO1.Journal of bacteriology · 2016Article
Corrections and comments
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Authors and funding
12 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe here a highly efficient procedure for conditional mutagenesis in Plasmodium. The procedure uses the site-specific recombination FLP-FRT system of yeast and targets the pre-erythrocytic stages of the rodent Plasmodium parasite P. berghei, including the sporozoite stage and the subsequent liver stage. The technique consists of replacing the gene under study by an FRTed copy (i.e., flanked by FRT sites) in the erythrocytic stages of a parasite clone that expresses the flip (FLP) recombinase stage-specifically--called the 'deleter' clone. We present the available deleter clones, which express FLP at different times of the parasite life cycle, as well as the schemes and tools for constructing new deleter parasites. We also outline and discuss the various strategies for exchanging a wild-type gene with an FRTed copy and for generating conditional gene knockout or knockdown parasite clones. Finally, we detail the protocol for obtaining sporozoites that lack a protein of interest and for monitoring sporozoite-specific DNA excision and depletion of the target protein. The protocol should allow the functional analysis of any essential protein in the sporozoite, liver stage or hepatic merozoite stages of rodent Plasmodium parasites.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.