ArticleMolecular and biochemical parasitology2009
Site-specific DNA double-strand breaks greatly increase stable transformation efficiency in Trypanosoma brucei.
Article in Molecular and biochemical parasitology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed, 44 citations in OpenAlex.
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- Paving the Way: Contributions of Big Data to Apicomplexan and Kinetoplastid Research.Frontiers in cellular and infection microbiology · 2022Review
- 4E Interacting Protein as a Potential Novel Drug Target for Nucleoside Analogues inMicroorganisms · 2021Article
- TbSAP is a novel chromatin protein repressing metacyclic variant surface glycoprotein expression sites in bloodstream form Trypanosoma brucei.Nucleic acids research · 2021Article
- Reliable, scalable functional genetics in bloodstream-form Trypanosoma congolense in vitro and in vivo.PLoS pathogens · 2021Article
- ATR Kinase Is a Crucial Player Mediating the DNA Damage Response inFrontiers in cell and developmental biology · 2020Article
- Persistent DNA Damage Foci and DNA Replication with a Broken Chromosome in the African Trypanosome.mBio · 2019Article
- A Seamless Gene Deletion Method and Its Application for Regulation of Higher Alcohols and Ester in BaijiuBioMed research international · 2019Article
- Inducible high-efficiency CRISPR-Cas9-targeted gene editing and precision base editing in African trypanosomes.Scientific reports · 2018Article
- Recruitment kinetics of the homologous recombination pathway in procyclic forms of Trypanosoma brucei after ionizing radiation treatment.Scientific reports · 2018Article
- CRISPR/Cas9-mediated gene modification and gene knock out in the human-infective parasite Trichomonas vaginalis.Scientific reports · 2018Article
- The Cytological Events and Molecular Control of Life Cycle Development of Trypanosoma brucei in the Mammalian Bloodstream.Pathogens (Basel, Switzerland) · 2017Review
- Homologous Recombination in Protozoan Parasites and Recombinase Inhibitors.Frontiers in microbiology · 2017Review
- Identification by Random Mutagenesis of Functional Domains in KREPB5 That Differentially Affect RNA Editing between Life Cycle Stages of Trypanosoma brucei.Molecular and cellular biology · 2015Article
- I-SceI-mediated double-strand DNA breaks stimulate efficient gene targeting in the industrial fungus Trichoderma reesei.Applied microbiology and biotechnology · 2015Article
- Genome-scale RNAi screens for high-throughput phenotyping in bloodstream-form African trypanosomes.Nature protocols · 2015Article
- High-efficiency genome editing and allele replacement in prototrophic and wild strains of Saccharomyces.Genetics · 2014Article
- Drug target identification using a trypanosome overexpression library.Antimicrobial agents and chemotherapy · 2014Article
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
Abstract
Genetic manipulation in African trypanosomes typically relies upon electroporation with chromosomal integration of DNA constructs by homologous recombination. Relatively little is known about chromosomal recombination and repair in these organisms however and low transformation efficiency and position effects can limit forward genetic approaches. In yeast and mammalian cells, site-specific DNA double-strand breaks (DSBs) stimulate targeted integration through homologous recombination-based repair where the exogenous DNA serves as the template. We have explored the effect of DSBs on targeted integration in bloodstream-form Trypanosoma brucei, focusing on the ribosomal RNA-spacer target commonly used to integrate recombinant constructs. DSB-repair within the ribosomal RNA tandem gene-repeats is likely dominated by single-strand annealing allowing approximately 80% of cells to survive the break. In the presence of exogenous DNA, transformation efficiency is increased approximately 250-fold by DSB-induction. In the example presented, more than 1% of cells that survive the procedure were transformed generating 80,000 transformants from a typical experiment.
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