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https://www.arca.fiocruz.br/handle/icict/52921
EFFICIENT CRISPR-CAS9-MEDIATED GENOME EDITING FOR CHARACTERIZATION OF ESSENTIAL GENES IN TRYPANOSOMA CRUZI
Author
Affilliation
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Illkirch, France. / Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Department of Integrated Structural Biology. Illkirch, France. / Loschmidt Laboratories. Department of Experimental Biology & RECETOX. Faculty of Science. Masaryk University. Brno, Czech Republic.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Programa de Pós-Graduação em Biologia Celular e Molecular. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Illkirch, France. / Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Department of Integrated Structural Biology. Illkirch, France.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Illkirch, France. / Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Department of Integrated Structural Biology. Illkirch, France. / Loschmidt Laboratories. Department of Experimental Biology & RECETOX. Faculty of Science. Masaryk University. Brno, Czech Republic.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Programa de Pós-Graduação em Biologia Celular e Molecular. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Illkirch, France. / Université de Strasbourg, CNRS, INSERM. Institut de Génétique et de Biologie Moléculaire et Cellulaire. Department of Integrated Structural Biology. Illkirch, France.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
Abstract
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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