Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi

This protocol outlines a new genetic complementation strategy to investigate gene function in Trypanosoma cruzi, the parasite causing Chagas disease. We combine CRISPR-Cas9 technology with recombination of variants of the target gene containing the desired mutations that are resistant to Cas9-cleavage, which enables detailed investigation of protein function. This experimental strategy overcomes some of the limitations associated with gene knockouts in T. cruzi. For complete details on the use and execution of this protocol, please refer to Marek et al. (2021).

Automated summary

This study presents a novel genetic complementation strategy for investigating gene function in Trypanosoma cruzi, the parasite responsible for Chagas disease. By combining CRISPR-Cas9 technology with recombination of resistant variants of the target gene containing desired mutations, detailed exploration of protein function is made possible. This experimental approach overcomes some of the limitations associated with traditional gene knockouts in T. cruzi.