Editing the Mouse Genome Using the CRISPR–Cas9 System

Adam Williams; Jorge Henao-Mejia; Richard A. Flavell

doi:10.1101/pdb.top087536

Editing the Mouse Genome Using the CRISPR–Cas9 System

¹The Jackson Laboratory for Genomic Medicine, Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, Connecticut 06032;
²Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104;
³Division of Transplant Immunology, Department of Pathology and Laboratory Medicine, Children’s Hospital of Philadelphia, University of Pennsylvania, Philadelphia, Pennsylvania 19104;
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

↵6 These authors contributed equally to this work.

Abstract

The ability to modify the murine genome is perhaps one of the most important developments in modern biology. However, traditional methods of genomic engineering are costly and relatively clumsy in their approach. The use of programmable nucleases such as zinc finger nucleases and transcription activator-like effector nucleases significantly improved the precision of genome-editing technology, but the design and use of these nucleases remains cumbersome and prohibitively expensive. The CRISPR–Cas9 system is the next installment in the line of programmable nucleases; it provides highly efficient and precise genome-editing capabilities using reagents that are simple to design and inexpensive to generate. Furthermore, with the CRISPR–Cas9 system, it is possible to move from a hypothesis to an in vivo mouse model in less than a month. The simplicity, cost effectiveness, and speed of the CRISPR–Cas9 system allows researchers to tackle questions that otherwise would not be technically or financially viable. In this introduction, we discuss practical considerations for the use of Cas9 in genome engineering in mice.