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-cleava...

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Detalles Bibliográficos
Autores principales: Picchi-Constante, Gisele Fernanda Assine, Hiraiwa, Priscila Mazzocchi, Marek, Martin, Rogerio, Vanessa Zulkievicz, Guerra-Slompo, Eloise Pavão, Romier, Christophe, Zanchin, Nilson Ivo Tonin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9048117/
https://www.ncbi.nlm.nih.gov/pubmed/35496799
http://dx.doi.org/10.1016/j.xpro.2022.101324
Descripción
Sumario: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).

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