Generation of Genome-Edited Mice by Cytoplasmic Injection of CRISPR-Cas9 RNA

Takuro Horii; Izuho Hatada

doi:10.1007/978-1-0716-3016-7_6

Generation of Genome-Edited Mice by Cytoplasmic Injection of CRISPR-Cas9 RNA

Methods Mol Biol. 2023:2637:75-86. doi: 10.1007/978-1-0716-3016-7_6.

Authors

Takuro Horii¹, Izuho Hatada^{2

3}

Affiliations

¹ Laboratory of Genome Science, Biosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Gunma, Japan. horii@gunma-u.ac.jp.
² Laboratory of Genome Science, Biosignal Genome Resource Center, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, Gunma, Japan. hatada@gunma-u.ac.jp.
³ Viral Vector Core, Gunma University Initiative for Advanced Research (GIAR), Maebashi, Gunma, Japan. hatada@gunma-u.ac.jp.

PMID: 36773139
DOI: 10.1007/978-1-0716-3016-7_6

Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) (CRISPR-Cas9) gene editing enables rapid production of genome-edited animals. The Cas9/guide RNA (gRNA) component can be introduced into zygotes in several ways. Here, we provide an instructional guide for the generation of knockout mice using cytoplasmic injection of in vitro transcribed Cas9 RNA and gRNA.

Keywords: CRISPR-Cas9; Cytoplasmic injection; In vitro transcription; Microinjection; Mouse.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Associated Protein 9 / genetics
CRISPR-Cas Systems* / genetics
Cytoplasm
Gene Editing*
Mice
Mice, Knockout
RNA, Guide, CRISPR-Cas Systems

Substances

CRISPR-Associated Protein 9
RNA, Guide, CRISPR-Cas Systems