Simple Protocol for Generating and Genotyping Genome‐Edited Mice With CRISPR‐Cas9 Reagents. Issue 1 (11th March 2020)

• Record Type: Journal Article
• Authors: Fernández, Almudena; Morín, Matías; Muñoz‐Santos, Diego; Josa, Santiago; Montero, Andrea; Rubio‐Fernández, Marcos; Cantero, Marta; Fernández, Julia; del Hierro, María Jesús; Castrillo, Marta; Moreno‐Pelayo, Miguel Ángel; Montoliu, Lluís
• Editors: Auwerx, Johan; Ackerman, Susan L.; Brown, Stephen D.; Justice, Monica J.; Nadeau, Joseph
• Is Part Of: Current protocols in mouse biology. Volume 10:Issue 1(2020)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-03-11
• Subjects: animal model -- CRISPR -- cytoplasmic injection and electroporation -- genome editing -- in vivo gene editing -- mouse model -- next‐generation sequencing; Mice -- Genetics -- Laboratory manuals; Mice; Animals, Laboratory; Research; Mice -- Genetics; Periodicals; Laboratory manuals; 616.027333
• DOI: 10.1002/cpmo.69
• Languages: English
• ISSNs: 2161-2617

Abstract

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