Generation of Genetically Modified Mice using the CRISPR-Cas9 Genome-Editing System

摘要

Genetically modified mice are extremely valuable tools for studying gene function and human diseases. Although the generation of mice with specific genetic modifications through traditional methods using homologous recombination in embryonic stem (ES) has been invaluable in the last two decades, it is an extremely costly, time-consuming and in some cases uncertain technology. The recently described CRISPR/Cas9 genome-editing technology significantly reduces the time and the cost that are required to generate genetically engineered mice, allowing scientist to test more precise and bold hypothesis in vivo. Using this revolutionary methodology we have generated more than one hundred novel genetically engineered mouse strains. In the current protocol, we describe in detail the optimal conditions to generate mice carrying point mutations, chromosomal deletions, conditional alleles, fusion tags or endogenous reporters.