CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae

Gene replacements through homologous recombination (HR) have been extensively used for functional genomic studies. However, the general efficiency of HR repair can be low in filamentous fungi and the process laborious. Here, we provide a detailed protocol for efficient gene editing by inserting dono...

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Detalles Bibliográficos
Autores principales: Huang, Jun, Cook, David E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8717591/
https://www.ncbi.nlm.nih.gov/pubmed/35005648
http://dx.doi.org/10.1016/j.xpro.2021.101072
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author Huang, Jun
Cook, David E.
author_facet Huang, Jun
Cook, David E.
author_sort Huang, Jun
collection PubMed
description Gene replacements through homologous recombination (HR) have been extensively used for functional genomic studies. However, the general efficiency of HR repair can be low in filamentous fungi and the process laborious. Here, we provide a detailed protocol for efficient gene editing by inserting donor DNA into a region of interest following Cas12a ribonucleoprotein (RNP)-mediated DNA double-strand break. We demonstrate this protocol using Magnaporthe oryzae (synonym of Pyricularia oryzae), a model plant pathogenic fungus that is used to study plant-fungal interactions. For complete details on the use and execution of this protocol, please refer to Huang et al. (2021).
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spelling pubmed-87175912022-01-06 CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae Huang, Jun Cook, David E. STAR Protoc Protocol Gene replacements through homologous recombination (HR) have been extensively used for functional genomic studies. However, the general efficiency of HR repair can be low in filamentous fungi and the process laborious. Here, we provide a detailed protocol for efficient gene editing by inserting donor DNA into a region of interest following Cas12a ribonucleoprotein (RNP)-mediated DNA double-strand break. We demonstrate this protocol using Magnaporthe oryzae (synonym of Pyricularia oryzae), a model plant pathogenic fungus that is used to study plant-fungal interactions. For complete details on the use and execution of this protocol, please refer to Huang et al. (2021). Elsevier 2021-12-24 /pmc/articles/PMC8717591/ /pubmed/35005648 http://dx.doi.org/10.1016/j.xpro.2021.101072 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Huang, Jun
Cook, David E.
CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae
title CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae
title_full CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae
title_fullStr CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae
title_full_unstemmed CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae
title_short CRISPR-Cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus Magnaporthe oryzae
title_sort crispr-cas12a ribonucleoprotein-mediated gene editing in the plant pathogenic fungus magnaporthe oryzae
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8717591/
https://www.ncbi.nlm.nih.gov/pubmed/35005648
http://dx.doi.org/10.1016/j.xpro.2021.101072
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AT cookdavide crisprcas12aribonucleoproteinmediatedgeneeditingintheplantpathogenicfungusmagnaportheoryzae

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