ArticleOpen biology2015
A toolkit enabling efficient, scalable and reproducible gene tagging in trypanosomatids.
Article in Open biology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 153 papers.
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Who cites it
153 citing papers in PubMed.
- Identification of a conserved gene family with an essential role inProceedings of the National Academy of Sciences of the United States of America · 2026Article
- Regulation of Leishmania surface coat proteins by the nuclear protein ESB1.Journal of cell science · 2026Article
- Proteomic Identification of Small-Subunit Ribosome Assembly Factors inJournal of proteome research · 2026Article
- Article
- Tubulin polyglutamylases TTLL4C and TTLL6B are essential for maintaining cytoskeletal integrity inmSphere · 2026Article
- The trypanosome mRNA decapping enzyme ALPH1 prefers caps without m7G methylation and produces diphosphate RNA.Nucleic acids research · 2026Article
- Article
- Trypanosome bloodstream-specific flagellum attachment proteins can mediate attachment in an insect surface coat environment.Journal of cell science · 2026Article
- Protein Localization with Streptavidin-Imaging: Boost Your Signal, Reduce Noise, Overcome Antibody Accessibility Problems, and Time-Travel to the Past.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Alkyl gallates disrupt Trypanosoma brucei lipid droplets.PloS one · 2026Article
- Effects of microtubule (de)tyrosination on the morphology and motility of Trypanosoma brucei and cross-talk with polyglutamylation.Biology open · 2025Article
- Visualization of Trypanosoma brucei flagellar pocket collar biogenesis identifies two new cytoskeletal structures.PLoS biology · 2025Article
- Next generation genetic screens in kinetoplastids.Nucleic acids research · 2025Review
- Infectiousness of Leishmania major to Phlebotomus papatasi: differences between natural reservoir host Meriones shawi and laboratory model BALB/c mice.PLoS neglected tropical diseases · 2025Article
- TblncRNA-23, a long non-coding RNA transcribed by RNA polymerase I, regulates developmental changes in Trypanosoma brucei.Nature communications · 2025Article
- Proteins with proximal-distal asymmetries in axoneme localisation control flagellum beat frequency.Nature communications · 2025Article
- A comprehensive toolkit for protein localization and functional analysis in trypanosomatids.Open biology · 2025Article
- Detailed characterisation of the trypanosome nuclear pore architecture reveals conserved asymmetrical functional hubs that drive mRNA export.PLoS biology · 2025Article
- A high-throughput protein tagging toolkit that retains endogenous untranslated regions for studying gene regulation in kinetoplastids.Open biology · 2025Article
- KRBP72 facilitates ATPase-dependent editing progression through a structural roadblock in mitochondrial A6 mRNA.Nucleic acids research · 2025Article
93 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
One of the first steps in understanding a protein's function is to determine its localization; however, the methods for localizing proteins in some systems have not kept pace with the developments in other fields, creating a bottleneck in the analysis of the large datasets that are generated in the post-genomic era. To address this, we developed tools for tagging proteins in trypanosomatids. We made a plasmid that, when coupled with long primer PCR, can be used to produce transgenes at their endogenous loci encoding proteins tagged at either terminus or within the protein coding sequence. This system can also be used to generate deletion mutants to investigate the function of different protein domains. We show that the length of homology required for successful integration precluded long primer PCR tagging in Leishmania mexicana. Hence, we developed plasmids and a fusion PCR approach to create gene tagging amplicons with sufficiently long homologous regions for targeted integration, suitable for use in trypanosomatids with less efficient homologous recombination than Trypanosoma brucei. Importantly, we have automated the primer design, developed universal PCR conditions and optimized the workflow to make this system reliable, efficient and scalable such that whole genome tagging is now an achievable goal.
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Registered trials
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.