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.