Simple Protocol for Generating and Genotyping Genome‐Edited Mice With CRISPR‐Cas9 Reagents

Abstract The simple protocol described in this article aims to provide all required information, as a comprehensive, easy‐to‐follow step‐by‐step method, to ensure the generation of the expected genome‐edited mice. Here, we provide protocols for the preparation of CRISPR‐Cas9 reagents for microinjection and electroporation into one‐cell mouse embryos to create knockout or knock‐in mouse models, and for genotyping the resulting offspring with the latest innovative next‐generation sequencing methods. © 2020 by John Wiley & Sons, Inc. Basic Protocol 1 : Designing the best RNA guide for your gene disruption/editing strategy Basic Protocol 2 : Preparing and validating CRISPR‐Cas9 reagents Basic Protocol 3 : Preparing and injecting CRISPR‐Cas9 compounds into fertilized mouse oocytes Basic Protocol 4 : Genotyping genome‐edited mice Support Protocol : Genotyping for CRISPR‐generated “indel” mutations