ArticleMolecular and biochemical parasitology2018
Optimising genetic transformation of Trypanosoma cruzi using hydroxyurea-induced cell-cycle synchronisation.
Article in Molecular and biochemical parasitology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 13 citations in OpenAlex.
- Remodeling of tRNA modification in Trypanosoma cruzi life forms.PLoS pathogens · 2026Article
- Knocking out histidine ammonia-lyase by using CRISPR-Cas9 abolishes histidine role in the bioenergetics and the life cycle ofMicrobial cell (Graz, Austria) · 2025Article
- Article
- Cell Synchronization Enhances Nuclear Transformation and Genome EditingACS synthetic biology · 2020Article
- Non-invasive monitoring of drug action: A new live in vitro assay design for Chagas' disease drug discovery.PLoS neglected tropical diseases · 2020Article
- Trypanocidal Mechanism of Action andInternational journal of molecular sciences · 2019Article
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Authors and funding
5 authors at 1 institution in 2 countries.
Funding
Abstract
The limited flexibility and time-consuming nature of the genetic manipulation procedures applicable to Trypanosoma cruzi continue to restrict the functional dissection of this parasite. We hypothesised that transformation efficiency could be enhanced if electroporation was timed to coincide with DNA replication. To test this, we generated epimastigote cultures enriched at the G1/S boundary using hydroxyurea-induced cell-cycle synchronisation, and then electroporated parasites at various time points after release from the cell-cycle block. We found a significant increase in transformation efficiency, with both episomal and integrative constructs, when cultures were electroporated 1 h after hydroxyurea removal. It was possible to generate genetically modified populations in less than 2 weeks, compared to the normal 4-6 weeks, with a 5 to 8-fold increase in the number of stably transformed clones. This straightforward optimisation step can be widely applied and should help streamline functional studies in T. cruzi.
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