Generation of Genetically Modified Mice Using the CRISPR–Cas9 Genome-Editing System
- Jorge Henao-Mejia1,2,6,7,
- Adam Williams3,6,7,
- Anthony Rongvaux4,
- Judith Stein4,5,
- Cynthia Hughes4,5 and
- Richard A. Flavell4,5,7
- 1Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104;
- 2Division of Transplant Immunology, Department of Pathology and Laboratory Medicine, Children’s Hospital of Philadelphia, University of Pennsylvania, Philadelphia, Pennsylvania 19104;
- 3The Jackson Laboratory for Genomic Medicine, Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, Connecticut 06032;
- 4Department of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520;
- 5Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut 06520
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↵6 These authors contributed equally to this work.
Abstract
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 cells 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 scientists to test more precise and bold hypotheses in vivo. Using this revolutionary methodology we have generated more than 100 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.
Footnotes
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↵7 Correspondence: jhena{at}mail.med.upenn.edu, adam.williams{at}jax.org, richard.flavell{at}yale.edu
- © 2016 Cold Spring Harbor Laboratory Press
