CRISPR/Cas9 Genome Editing

The Transgenic Animal Facility has been using CRISPR/Cas9 technology to create gene-edits in mice since 2014.  We are one of the elite facilities within the United States to offer a comprehensive array of services from design to synthesis to generation of genome-edited animal models now including mice, rats and swine.

Instead of 1.5-2 years to make a genome edited mouse, it now takes as little as 4 months! 

Some examples of modifications TAF has successfully created:

  • Gene knockout
  • Precise deletion
  • Small knock-in (amino acid, tag)
  • Floxed allele
  • Reporter or Cre knock-in 

Designing your Genome-edit

Dustin Rubinstein runs the Translational Genomics Facility within the Biotechnology Center and will assist you in designing your CRISPR guides and obtaining injection-ready reagents. Contact him at crubinstein@wisc.edu

Microinjection of embryos to produce gene-edits in mice and rats

After a purified gRNA has been created by the TGF, an injection day will be scheduled with Kathy Krentz in the TAF. The gRNA, Cas9, ssODN, and/or plasmid DNA (if applicable) will be microinjected into fertilized one-cell embryos. The injected one-cell embryos are then implanted into the pseudopregnant recipients and offspring are born 19 days (mice) or 22 days (rats) later. At weaning, animals are genotyped to detect presence of edits.  Founders can be bred within the Biotechnology Center or shipped to the investigators for breeding.