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.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.