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

Gisele Fernanda Assine Picchi-Constante , Priscila Mazzocchi Hiraiwa , Martin Marek (1, 2) , Vanessa Zulkievicz Rogerio , Eloise Pavão Guerra-Slompo , Christophe Romier (2, 1) , Nilson Ivo Tonin Zanchin

1 IGBMC - Institut de génétique et biologie moléculaire et cellulaire
2 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire

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).

Domaines: Génétique

Gisele Fernanda Assine Picchi-Constante, Priscila Mazzocchi Hiraiwa, Martin Marek, Vanessa Zulkievicz Rogerio, Eloise Pavão Guerra-Slompo, et al.. Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi. STAR Protocols, 2022, 3 (2), ⟨10.1016/j.xpro.2022.101324⟩. ⟨hal-04219152⟩