Genome Editing of Medaka

Rie Hara; Satoshi Ansai; Masato Kinoshita

doi:10.1007/978-1-0716-3016-7_25

Genome Editing of Medaka

Methods Mol Biol. 2023:2637:325-339. doi: 10.1007/978-1-0716-3016-7_25.

Authors

Rie Hara¹, Satoshi Ansai², Masato Kinoshita³

Affiliations

¹ Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
² Graduate School of Life Sciences, Tohoku University, Miyagi, Japan.
³ Graduate School of Agriculture, Kyoto University, Kyoto, Japan. kinoshita.masato.4e@kyoto-u.ac.jp.

PMID: 36773158
DOI: 10.1007/978-1-0716-3016-7_25

Abstract

Medaka (Oryzias latipes), along with zebrafish (Danio rerio), is a useful experimental model fish. Here, we describe a simple method for generating medaka gene knockout strains using an automated microchip electrophoresis system. We also describe a method for targeted gene knockin using a plasmid carrying a sequence that does not cause off-target effects in medaka. Additionally, knockin method without plasmid cloning is described.

Keywords: CRISPR/Cas; Genome editing; Heteroduplex mobility assay; Medaka; Targeted integration; Targeted mutagenesis.

MeSH terms

Animals
Gene Editing* / methods
Gene Knockout Techniques
Oryzias* / genetics
Plasmids / genetics
Zebrafish / genetics