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
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author Picchi-Constante, Gisele Fernanda Assine
Hiraiwa, Priscila Mazzocchi
Marek, Martin
Rogerio, Vanessa Zulkievicz
Guerra-Slompo, Eloise Pavão
Romier, Christophe
Zanchin, Nilson Ivo Tonin
author_facet Picchi-Constante, Gisele Fernanda Assine
Hiraiwa, Priscila Mazzocchi
Marek, Martin
Rogerio, Vanessa Zulkievicz
Guerra-Slompo, Eloise Pavão
Romier, Christophe
Zanchin, Nilson Ivo Tonin
author_sort Picchi-Constante, Gisele Fernanda Assine
collection PubMed
description 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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spelling pubmed-90481172022-04-29 Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi Picchi-Constante, Gisele Fernanda Assine Hiraiwa, Priscila Mazzocchi Marek, Martin Rogerio, Vanessa Zulkievicz Guerra-Slompo, Eloise Pavão Romier, Christophe Zanchin, Nilson Ivo Tonin STAR Protoc Protocol 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). Elsevier 2022-04-21 /pmc/articles/PMC9048117/ /pubmed/35496799 http://dx.doi.org/10.1016/j.xpro.2022.101324 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Picchi-Constante, Gisele Fernanda Assine
Hiraiwa, Priscila Mazzocchi
Marek, Martin
Rogerio, Vanessa Zulkievicz
Guerra-Slompo, Eloise Pavão
Romier, Christophe
Zanchin, Nilson Ivo Tonin
Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_full Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_fullStr Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_full_unstemmed Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_short Efficient CRISPR-Cas9-mediated genome editing for characterization of essential genes in Trypanosoma cruzi
title_sort efficient crispr-cas9-mediated genome editing for characterization of essential genes in trypanosoma cruzi
topic Protocol
url 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
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