Cas9-mediated-KO

The generation of a Cas9-mediated KO can be achieved by the injection of Cas9 protein or mRNA together with a guide RNA (gRNA) that determines the specificity, into a mouse zygote. The enzyme will introduce DNA breaks at the location predetermined by the co-injected gRNA. These breaks are repaired by the sometimes faulty non-homologous end joining repair mechanism (NHEJ), leading in a large number of founder animals to random insertions and deletions. 

This service offers the generation of knockout animals by pronuclear injection of components of the CRISPR/Cas9 system. Injections are routinely performed in C57BL6/J background, with other backgrounds possible by request (price may vary). The embryo transfers following microinjections are performed under SPF conditions, thus allowing founders to be introduced into most facilities. Please contact us directly for special requests such as injections in other mouse strains.

1. A completely filled out Request Form
2. The GMO datasheet as requested by the veterinary office
3. An iRATS breeding project opened under researchers own license to be used for weaned offspring (iRATS users only)
4. The genebank (*.gbk or *.gb) file with the sequence of interest marking the KO region.
5. 10 μg of a PCR product spanning the KO region at > 150 ng/μl. 

The cost of this service is 5800 CHF for inbred (C57BL/6J) mouse strains. This fee covers the donor animals, the injection of CRISPR/Cas9 components, embryo transfers, collection of biopsies and subsequent animal care costs until the founders are turned over to the researcher. An additional fee of 700 CHF will be charged when founders should already be bred to wt animals, before transfer to the TCF.

For terms and conditions please click  here.

In case of questions, please contact Johannes vom Berg before submitting a microinjection request. Every Friday we offer a “Targeting Club” at 9:00 am in our seminar rooms (Wagistr. 12, Schlieren or Sternwartstrasse 6, Unispital, Zürich), where you are welcome to discuss your project with us (see also upcoming dates).