@inbook{06653680c71e48cbb314f11c70575105,
title = "Genome Editing Using CRISPR/Cas9 System in the Rice Blast Fungus",
abstract = "Fast and flexible genome manipulation is a powerful strategy for an in-depth understanding of molecular mechanisms in biological research. In recent years, CRISPR/Cas9-mediated genome editing has been used as a reliable genome manipulation method in a broad range of biological research including studies of filamentous fungi. The CRISPR/Cas9 system comprises a single-guide RNA (sgRNA) and a Cas9 protein, and the Cas9/sgRNA complex catalyzes a DNA double-strand break at the desired genomic locus. This protocol describes a fundamental CRISPR/Cas9 methodology that includes the design of the target sequence, construction of the CRISPR/Cas9 expression vector, and transformation for genome editing in Pyricularia (Magnaporthe) oryzae. This allows efficient targeted gene disruption, base editing, and reporter gene knock-in without any additional modifications of the host components. This protocol would be suitable for applying other CRISPR/Cas technologies and various functional genomics in P. oryzae.",
keywords = "CRISPR/Cas9, Filamentous fungi, Functional genomics, Genome editing, Homologous recombination, Pyricularia (Magnaporthe) oryzae",
author = "Takayuki Arazoe",
note = "Funding Information: This work is supported by the Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research (A) (Grant Number 16H02536) and Grant-in-Aid for Young Scientists (B) (Grant Number 15 K18647). Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2021",
doi = "10.1007/978-1-0716-1613-0_12",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "149--160",
booktitle = "Methods in Molecular Biology",
}