期刊
STAR PROTOCOLS
卷 3, 期 1, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.xpro.2021.101072
关键词
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资金
- United State Department of Agriculture-National Institute of Food and Agriculture (USDA-NIFA) [1936800]
- National Science Foundation Division of Molecular and Cellular Biosciences-Systems and Synthetic Biology
- [2018-67013- 28492]
This article presents 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. The protocol is demonstrated using Magnaporthe oryzae, a model plant pathogenic fungus used in the study of plant-fungal interactions.
Gene replacements through homologous recombination (HR) have been exten-sively 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 or-yzae (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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