ArticleMolecular and biochemical parasitology2008
CRE recombinase-based positive-negative selection systems for genetic manipulation in Trypanosoma brucei.
Article in Molecular and biochemical parasitology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 42 citations in OpenAlex.
- Article
- KREPA6 functions in RNA editing catalytic complex structural organization and gRNA utilization in Trypanosoma brucei.Nucleic acids research · 2026Article
- SLAM-seq reveals independent contributions of RNA processing and stability to gene expression in African trypanosomes.Nucleic acids research · 2025Article
- Generation of a bloodstream form Trypanosoma brucei double glycosyltransferase null mutant competent in receptor-mediated endocytosis of transferrin.PLoS pathogens · 2024Article
- POLIE suppresses telomerase-mediated telomere G-strand extension and helps ensure proper telomere C-strand synthesis in trypanosomes.Nucleic acids research · 2022Article
- Distinct roles for H4 and H2A.Z acetylation in RNA transcription in African trypanosomes.Nature communications · 2020Article
- Trypanosoma brucei RAP1 Has Essential Functional Domains That Are Required for Different Protein Interactions.mSphere · 2020Article
- Article
- Article
- Exploiting CRISPR-Cas9 technology to investigate individual histone modifications.Nucleic acids research · 2018Article
- A dual regulatory circuit consisting of S-adenosylmethionine decarboxylase protein and its reaction product controls expression of the paralogous activator prozyme in Trypanosoma brucei.PLoS pathogens · 2018Article
- GT-rich promoters can drive RNA pol II transcription and deposition of H2A.Z in African trypanosomes.The EMBO journal · 2017Article
- In vivo cleavage specificity of Trypanosoma brucei editosome endonucleases.Nucleic acids research · 2017Article
- Differential Editosome Protein Function between Life Cycle Stages of Trypanosoma brucei.The Journal of biological chemistry · 2015Article
- Suppression of subtelomeric VSG switching by Trypanosoma brucei TRF requires its TTAGGG repeat-binding activity.Nucleic acids research · 2014Article
- Trypanosoma brucei (UMP synthase null mutants) are avirulent in mice, but recover virulence upon prolonged culture in vitro while retaining pyrimidine auxotrophy.Molecular microbiology · 2013Article
- Strategies to construct null and conditional null Trypanosoma brucei mutants using Cre-recombinase and loxP.Molecular and biochemical parasitology · 2013Article
- Review
- Pyrimidine biosynthesis is not an essential function for Trypanosoma brucei bloodstream forms.PloS one · 2013Article
- Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
The limited repertoire of drug-resistance markers imposes a serious obstacle to genetic manipulation of Trypanosoma brucei. Here we describe experiments with a fusion protein that allows positive selection for genome integration followed by CRE recombinase-mediated excision of the marker cassette that can be selected by ganciclovir, although the excision event is so efficient that selection is not strictly necessary. We describe two variants of the tetracycline-inducible pLEW100-based CRE-expression vector that reduced its toxicity when stably integrated into the genome, and we demonstrate that transient transfection of circular pLEW100-CRE is highly efficient at catalyzing marker excision. We used this approach to delete the last two enzymes of the pyrimidine synthesis pathway, creating a cell line that is resistant to fluoroorotic acid, which would allow the same enzymes (PYR6-5) to be used as an alternative negative selectable marker.
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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.